Team:NCTU Formosa/notes

From 2013.igem.org

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===About the notes===
===About the notes===
-
(tell something if there's anything to say.)
+
Our notes are the abbreviations of the experimental process, including titles, genes we transformed, plasmids we used, and resistances on the plasmids. We categorized these notes by date. Moreover, team members followed the iGEM protocol in each experimental step in order to get better experimental efficiency and the results we had expected.
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<h3>March 2013</h3>
+
===Material and Methods===
 +
====Minipreps of Plasmid DNA====
 +
We use Plasmid miniPREP kit (Genedirex Inc.) to obtain plasmid DNA for analysis, sequencing and cloning. Single colony is picked from LB agar plate and inoculated into LB medium with suitable antibiotics. Bacteria cells are harvest by centrifuge after incubation for 12 – 16 hours. Afterward, alkaline lysis is performed to release inside components, and DNA is purified by silicone resin-based column. The quality and amount of plasmid DNA is estimated by agarose gel electrophoresis stained by SyBr Safe (Invitrogen).
 +
 
 +
====BioBrick assembly====
 +
To digest the desired DNA, appropriate restriction enzymes and reaction buffer (all perched from New England Biolabs) is used in 20 μL reaction volume. Upstream BioBrick is digested with EcoRI-HF and SpeI in CutSmart buffer. Downstream BioBrick and backbone part are digested with XbaI and PstI, EcoRI-HF and PstI respectively in NEBuffer 2. Reactions take place in 37 ℃ water bath for 2 hours or overnight.
 +
 
 +
Ligation is performed using BioBrick Assembly Kit Ligation Protocol. All the reagent and enzymes are perched from New England Biolabs). Reactions take place in 37 ℃ water bath for 2 hours or overnight in 16 ℃ incubator.
 +
 
 +
====Transformation====
 +
DH5α ''E. coli'' competent cells (ECOS<sup>TM</sup> 101, Yeastern Biotech Co., Ltd.) were used for the propagation of plasmid DNA. Standard transformation protocol is used: First place 2 μL plasmid or 10μL product into 1.5 mL mini centrifuge tube and chilled on ice. Competent cells are melt on ice before transformation. After adding 33 – 50 μL competent cells, heat shock at 42 ℃ for 1 minutes. Place cells into LB medium to recover then plating on LB agar plate and incubate for 16 – 18 hours.
 +
 
 +
====PCR reaction====
 +
Taq DNA Polymerase 2x Master Mix Red (Ampliqon) and VF2 and VR primers (BBa_G00100 and BBa_G00101 respectively) are used for colony PCR. Single colony is picked and place into 10 μL PCR reagent, and followed by PCR with condition described below:
 +
*Predenature: 95 ℃，10 min
 +
*Denature, annealing and extension: [98 ℃, 10 s;  55 ℃, 30 s; 72 ℃, 1 m/kb + 30 s] × 25 cycles
 +
*Hold: 4 ℃
 +
 
 +
====Reagent and Medium====
 +
Lysogeny broth (LB): 10 g/L tryptone, 5 g/L yeast extract, 10 g/L NaCl with with suitable antibiotics.
 +
 
 +
LB agar plate: LB medium with 1.5% (w/v) agar.
 +
 
 +
50X TAE buffer: tris base 242 g/L, glacial acetic acid 57.1 mL/L, 0.05 M EDTA, pH 8.0
 +
Antibiotics working concentration:
 +
*Kanamycin      : 25 μg/mL
 +
*Ampicillin    : 100 μg/mL
 +
*Chloramphenicol: 20 μg/mL
 +
 
 +
 
 +
 
 +
</div>
 +
</div>
 +
<div class="li"><div class="card">
 +
 
 +
===March 2013===
 +
<html>
<!---------------------------------------- cal starts ---------------------------------------->
<!---------------------------------------- cal starts ---------------------------------------->
<div class="calendar">
<div class="calendar">
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<div class="daysmonth">
<div class="daysmonth">
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week">
+
<div class="week mweek">
<div class="day">
<div class="day">
-
<div class="daybar"><p>11</p></div>
+
<div class="daybar"><p>24</p></div>
-
<div class="dots">
+
-
</div>
+
-
 
+
-
<div class="open">
+
-
</div>
+
-
 
+
-
</div>
+
-
<div class="day">
+
-
<div class="daybar"><p>12</p></div>
+
-
<div class="dots">
+
-
</div>
+
-
 
+
-
<div class="open">
+
-
</div>
+
-
 
+
-
</div>
+
-
<div class="day">
+
-
<div class="daybar"><p>13</p></div>
+
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>mini-prep of cultivated AlsS, pSB1K3, pSB1A3, and pSB1C3 E.coli  (G2)</p></li>
 
</ul>
</ul>
-
</div>
 
-
 
-
</div>
 
-
<div class="day">
 
-
<div class="daybar"><p>14</p></div>
 
-
<div class="dots">
 
-
<ul>
 
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
-
</ul>
 
-
</div>
 
-
 
-
<div class="open">
 
-
<ul>
 
-
<li class="green e2"><p>digestion : [Zif268] ES &amp; [AlsS] XP &amp; [pSB1C3] EP  (G2)</p></li>
 
-
<li class="green e2"><p>ligation : insert[Zif268] ES &amp; [AlsS] XP/Vector[pSB1C3] EP  (G2)</p></li>
 
-
</ul>
 
-
</div>
 
-
 
-
</div>
 
-
<div class="day">
 
-
<div class="daybar"><p>15</p></div>
 
-
<div class="dots">
 
-
<ul>
 
-
</ul>
 
-
</div>
 
-
 
-
<div class="open">
 
-
<ul>
 
-
</ul>
 
-
</div>
 
-
 
-
</div>
 
-
<div class="day">
 
-
<div class="daybar"><p>16</p></div>
 
-
<div class="dots">
 
-
<ul>
 
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
-
</ul>
 
-
</div>
 
-
 
-
<div class="open">
 
-
<ul>
 
-
<li class="green e2"><p>Transformation of B0034</p></li>
 
-
<li class="green e2"><p>Cultivation of transformed E.coli on LBA plate</p></li>
 
-
</ul>
 
-
</div>
 
-
 
-
</div>
 
-
<div class="day brn">
 
-
<div class="daybar"><p>17</p></div>
 
-
<div class="dots">
 
-
</div>
 
-
 
-
<div class="open">
 
-
</div>
 
-
 
-
</div>
 
-
</div>
 
-
<!---------------------------------------- week start ---------------------------------------->
 
-
<div class="week">
 
-
<div class="day">
 
-
<div class="daybar"><p>18</p></div>
 
-
<div class="dots">
 
-
</div>
 
-
 
-
<div class="open">
 
</div>
</div>
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>19</p></div>
+
<div class="daybar"><p>25</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Transformation of Zif268+AlsS+pSB1C3 in DH5-alpha  (G2)</p></li>
 
-
<li class="green e2"><p>Point mutation HivC (PCR, DPN1 digest, TA cloning, transform) (G2)</p></li>
 
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>20</p></div>
+
<div class="daybar"><p>26</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of point mutation HivC  (G2)</p></li>
 
-
<li class="green e2"><p>PCR of insert fragment [Zif268+AlsS]  (G2)</p></li>
 
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>21</p></div>
+
<div class="daybar"><p>27</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>electrophoresis of insert fragment [Zif268+AlsS]----NOT OK  (G2)</p></li>
 
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>22</p></div>
+
<div class="daybar"><p>28</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Single colony isolation from pLac LBA plate, and cultivation of pLac E.coli in liquid LBA</p></li>
 
-
<li class="green e2"><p>Single colony isolation from pSB1K3 LBK plate, and cultivation of pSB1K3 E.coli in liquid LBK</p></li>
 
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>23</p></div>
+
<div class="daybar"><p>29</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>mini-prep of cultivated pLac &amp; pSB1K3 E.coli</p></li>
+
<li class="green e2"><p>Transformation of ho1(Kr), Cph8(Cr), 32℃ RBS(Ar) and cultivation  on three LB plates.</p></li>
 +
<li class="green e2"><p>transformation of PompC and
 +
cultivation  on LB-A plate</p></li>
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>24</p></div>
+
<div class="daybar"><p>30</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>digestion : [pSB1K3] EP</p></li>
+
<li class="green e3"><p>Single colony isolation from three plates and cultivation them in liquid LB</p></li>
 +
<li class="green e3"><p>Single colony isolation from PompC LB-C plate and cultivation in liquid LB-A</p></li>
 +
<li class="green e3"><p>mini-prep of cultivated PompC E.coli</p></li>
</ul>
</ul>
</div>
</div>
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</div>
</div>
</div>
</div>
 +
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week">
+
<div class="week mweek">
<div class="day">
<div class="day">
-
<div class="daybar"><p>25</p></div>
+
<div class="daybar"><p>31</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>Transformation of RBS  B0030+PSB1A3&B0034+PSB1A3 and cultivation on LB-A plate.</p></li>
 +
<li class="green e4"><p>Mini-prep for cultivated  ho1(Kr), Cph8(Cr), 32℃ RBS(Ar) PCR of ho1(Kr), Cph8(Cr), 32℃ RBS(Ar).</p></li>
 +
<li class="green e4"><p>Electrophoresis of ho1(Kr), Cph8(Cr), 32℃ RBS(Ar) ──NOT OK.</p></li>
 +
<li class="green e4"><p>PCR  of insert fragment
 +
[PompC+psB1A2]</p></li>
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>26</p></div>
+
<div class="daybar"><p></p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>27</p></div>
+
<div class="daybar"><p></p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>28</p></div>
+
<div class="daybar"><p></p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>29</p></div>
+
<div class="daybar"><p></p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Single colony isolation from Zif268 LBA plate, and cultivation of Zif268 E.coli in liquid LBA</p></li>
 
-
<li class="green e2"><p>Single colony isolation from AlsS LBA plate, and cultivation of AlsS E.coli in liquid LBA</p></li>
 
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>30</p></div>
+
<div class="daybar"><p></p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>mini-prep of cultivated Zif268 Ecoli &amp; AlsS Ecoli</p></li>
 
-
<li class="green e2"><p>digestion :[Zif268] ES/[AlsS] XP</p></li>
 
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>31</p></div>
+
<div class="daybar"><p></p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
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</div>
</div>
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week end ---------------------------------------->
-
</div>
 
</div>
</div>
</div>
</div>
 +
</div>
<!---------------------------------------- cal ends ---------------------------------------->
<!---------------------------------------- cal ends ---------------------------------------->
</div>
</div>
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<div class="week">
<div class="week">
<div class="day">
<div class="day">
-
<div class="daybar"><p>1</p></div>
+
<div class="daybar"><p></p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>ligation : insert Zif268 [ES] &amp; AlsS [XP]/Vector pSB1K3 [EP]</p></li>
 
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>2</p></div>
+
<div class="daybar"><p>1</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of Zif268+AlsS+pSB1K3 and cultivation on LB-K plate</p></li>
+
<li class="green"><p>Cultivation of B0030&B0034 Ar  E.coli  in liquid LB-A tubes.</p></li>
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>3</p></div>
+
<div class="daybar"><p>2</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green"><p>Mini-prep of cultivated B0030&B0034 E.coli and then digestion: [B0030]XP&[B0034]XP.</p></li>
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>4</p></div>
+
<div class="daybar"><p>3</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>digestion : [pSB1K3] EP</p></li>
 
-
<li class="green e2"><p>digestion : [pSB1K3] EP</p></li>
 
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>5</p></div>
+
<div class="daybar"><p>4</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
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</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>6</p></div>
+
<div class="daybar"><p>5</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
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</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>7</p></div>
+
<div class="daybar"><p>6</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e4"><p>PCR of insert fragment [Zif268+AlsS] OK</p></li>
+
<li class="green e2"><p>Electrophoresis of [B0030]XP&[B0034]XP Not OK.</p></li>
-
<li class="green e4"><p>DNA Sequencing OK</p></li>
+
<li class="green e2"><p>PCR of ho1 to check ho1  and transform to test the resistance.</p></li>
-
<li class="green e4"><p>ligation : point mutation HivC</p></li>
+
-
<li class="green e4"><p>TA cloning : point mutation HivC  </p></li>
+
</ul>
</ul>
</div>
</div>
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<div class="week">
<div class="week">
<div class="day">
<div class="day">
-
<div class="daybar"><p>8</p></div>
+
<div class="daybar"><p>7</p></div>
<div class="dots">
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
</ul>
</div>
</div>
<div class="open">
<div class="open">
 +
<li class="green e4"><p>Electrophoresis of [B0030]mini&dig XP and [B0034]mini&dig XP OK</p></li>
 +
<li class="green e4"><p>Cultivation of  B0030&B0034 Ar  E.coli in liquid LB-A tubes.</p></li>
 +
<li class="green e4"><p>Cultivation of ho1 in liquid LB-K and LB-K  plate</p></li>
 +
<li class="green e4"><p>Cultivation of Cph8+RBS in liquid LB-C.</p></li>
</div>
</div>
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>9</p></div>
+
<div class="daybar"><p>8</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
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<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>Single colony isolation from Zif268+AlsS LB-K plate, and cultivation of in liquid LB-K</p></li>
+
<li class="green"><p>Mini-prep of cultivated B0030&B0034 Ar E.coli. Mini-prep of ho1 & Cph8+RBS.</p></li>
</ul>
</ul>
</div>
</div>
Line 489: Line 434:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>10</p></div>
+
<div class="daybar"><p>9</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 500: Line 443:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>mini-prep of cultivated Zif268+ AlsS E. coli</p></li>
+
<li class="green"><p>Electrophoresis of mini ho1&Cph8+RBS.</p></li>
-
<li class="green e3"><p>transformation of point mutation HivC and cultivation on LB-A plate</p></li>
+
-
<li class="green e3"><p>transformation of B0034 and cultivation on LB-A plate</p></li>
+
</ul>
</ul>
</div>
</div>
Line 508: Line 449:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>11</p></div>
+
<div class="daybar"><p>10</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
Line 521: Line 462:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>12</p></div>
+
<div class="daybar"><p>11</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 531: Line 473:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Single colony isolation from HivC LB-A plate, and in liquid LB-A</p></li>
+
<li class="green e3"><p>PCR of mini ho1&Cph8+RBS</p></li>
-
<li class="green e2"><p>Single colony isolation from B0034 LB-A plate, and in liquid LB-A</p></li>
+
<li class="green e3"><p>Electrophoresis of ho1&Cph8+RBS(After PCR)</p></li>
 +
<li class="green e3"><p>Digestion: [ho1]EP&[Cph8+RBS]EP.</p></li>
</ul>
</ul>
</div>
</div>
Line 538: Line 481:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>13</p></div>
+
<div class="daybar"><p>12</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 547: Line 491:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>mini-prep of cultivated point mutation HivC E. coli &amp; B0034 E. coli </p></li>
+
<li class="green e2"><p>Transformation of Plac&Ptet& pcyA but there is no colony appearing in pcyA plate.</p></li>
 +
<li class="green e2"><p>Electrophoresis of digested ho1&Cph8+RBS (NOT OK).</p></li>
</ul>
</ul>
</div>
</div>
Line 553: Line 498:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>14</p></div>
+
<div class="daybar"><p>13</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 562: Line 506:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>digestion : Zif268+AlsS [XP]</p></li>
 
</ul>
</ul>
</div>
</div>
Line 571: Line 514:
<div class="week mweek">
<div class="week mweek">
<div class="day">
<div class="day">
-
<div class="daybar"><p>15</p></div>
+
<div class="daybar"><p>14</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 579: Line 524:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e2"><p>Cultivation of Ptet&Plac E.coli in LB tubes.</p></li>
 +
<li class="green e2"><p>Transformation: LuxR,B0015,J61048 and cultivation on LB-A plate</p></li>
</ul>
</ul>
</div>
</div>
Line 584: Line 531:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>16</p></div>
+
<div class="daybar"><p>15</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 592: Line 543:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>Mini-prep of cultivated Ptet&Plac E.coli</p></li>
 +
<li class="green e4"><p>Digestion:[Ptet]ES&[Plac]ES
 +
PCR of insert fragment pcyA</p></li>
 +
<li class="green e4"><p>Electrophoresis of [Ptet]dig ES&[pcyA]PCR&[Plac]dig ES.</p></li>
 +
<li class="green e4"><p>Single colony  isolation from J61048,B0015,LuxR  plate and cultivation  in  liquid LB-A mini-prep of cultivated LuxR,J61048,B0015</p></li>
</ul>
</ul>
</div>
</div>
Line 597: Line 553:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>17</p></div>
+
<div class="daybar"><p>16</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 606: Line 565:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>Single colony isolation from ilvD LB-A plate, and cultivation in liquid LB-A </p></li>
+
<li class="green e4"><p>PCR of mini Cph8+RBS</p></li>
 +
<li class="green e4"><p>Electrophoresis of Cph8+RBS(After PCR)──NOT OK.</p></li>
 +
<li class="green e4"><p>digestion: [J61048]EP,[B0015]EP,[LuxR]EP</p></li>
 +
<li class="green e4"><p>Electrophoresis of ter. mini [J61048] dig E/EP and t er. mini [B0015] dig E/EP and luxR mini dig E/EP</p></li>
</ul>
</ul>
</div>
</div>
Line 612: Line 574:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>18</p></div>
+
<div class="daybar"><p>17</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
Line 623: Line 585:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>mini-prep of cultivated ilvD E. coli</p></li>
+
<li class="green e3"><p>PCR of mini Cph8+RBS
-
<li class="green e3"><p>digestion : ilvD [EP] &amp; pSB1K3 [EP]</p></li>
+
Electrophoresis of Cph8+RBS(After PCR)──NOT OK.</p></li>
-
<li class="green e3"><p>transformation of 37℃ RBS and cultivation on LB-C plate</p></li>
+
<li class="green e3"><p>Electrophoresis of the products of digestion of ter. [J61048], ter. [B0015] , luxR</p></li>
 +
<li class="green e3"><p>transformation of tetR plasmid and cultivation on LB –A plate 3. Digestion: ter. [J61048], ter. [B0015] dig EP</p></li>
</ul>
</ul>
</div>
</div>
Line 631: Line 594:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>19</p></div>
+
<div class="daybar"><p>18</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 640: Line 602:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>Single colony isolation from HivC LB-A plate, and cultivation in liquid LB-A </p></li>
 
</ul>
</ul>
</div>
</div>
Line 646: Line 607:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>20</p></div>
+
<div class="daybar"><p>19</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
Line 658: Line 619:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e4"><p>digestion : B0034 [SP]</p></li>
+
<li class="green e4"><p>Cultivation of pcyA Kr E.coli on LB-K plates</li>
-
<li class="green e4"><p>gel extraction</p></li>
+
<li class="green e4"><p>mini-prep of cultivated tetR  E.coli</p></li>
-
<li class="green e4"><p>ligation :insert Zif268+AlsS [XP]/Vector B0034 [SP]</p></li>
+
<li class="green e4"><p>digestion: tetR+pSB1A2 dig EP</p></li>
-
<li class="green e4"><p>mini-prep of cultivated Hivc E. coli</p></li>
+
<li class="green e4"><p>electrophoresis of tetR dig EP,ter.[J61048 ] dig EP and ter.[B0015] dig EP</p></li>
</ul>
</ul>
</div>
</div>
Line 667: Line 628:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>21</p></div>
+
<div class="daybar"><p>20</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 677: Line 639:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>transformation of B0034+Zif268+AlsS and cultivation on LB-A plate</p></li>
+
<li class="green e3"><p>Cultivation of pcyA Kr E.coli in liquid LB-K tubes.</p></li>
-
<li class="green e2"><p>transformation of HivC and cultivation on LB-A plate</p></li>
+
<li class="green e3"><p>PCR of tetR,ter.[J61048] and ter.[B0015 ]mini</p></li>
 +
<li class="green e3"><p>electrophoresis of PCR products of tetR mini,J61048 mini and B0015 mini</p></li>
</ul>
</ul>
</div>
</div>
Line 688: Line 651:
<div class="week bigwk">
<div class="week bigwk">
<div class="day">
<div class="day">
-
<div class="daybar"><p>22</p></div>
+
<div class="daybar"><p>21</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
Line 704: Line 667:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e8"><p>PCR of insert fragment [B0034+Zif268+AlsS] OK</p></li>
+
<li class="green e8"><p>Single colony isolation of pcyA Kr E.coli</p></li>
-
<li class="green e8"><p>DNA Sequencing NOT OK</p></li>
+
<li class="green e8"><p>Mini-prep of cultivated pcyA Kr E.coli</p></li>
-
<li class="green e8"><p>digestion : B0034 [SP] &amp; Zif268+AlsS [XP]</p></li>
+
<li class="green e8"><p>Digestion:[pcyA]ES</p></li>
-
<li class="green e8"><p>gel extraction</p></li>
+
<li class="green e8"><p>Electrophoresis of [pcyA]mini&dig ES OK</p></li>
-
<li class="green e8"><p>ligation :insert Zif268+AlsS [XP]/Vector B0034+pSB1A3 [SP]</p></li>
+
<li class="green e8"><p>PCR of tetR,ter.[J61048] and ter.[B0015 ]mini</p></li>
-
<li class="green e8"><p>transformation of B0034+Zif268+AlsS and cultivation on LB-A plate</p></li>
+
<li class="green e8"><p>electrophoresis of PCR products of tetR mini,J61048 mini and B0015 mini</p></li>
-
<li class="green e8"><p>Single colony isolation from B0034 LB-A plate, and cultivation in liquid LB-A</p></li>
+
<li class="green e8"><p>electrophoresis of the digestion products of tetR dig EP and B0015 dig EP</p></li>
-
<li class="green e8"><p>SCI from ilvD LB-K plate, and cultivation in liquid LB-K</p></li>
+
<li class="green e8"><p>aliquot every 20ul of PompC,J61048 and LuxR</p></li>
</ul>
</ul>
</div>
</div>
Line 717: Line 680:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>23</p></div>
+
<div class="daybar"><p>22</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 729: Line 688:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e4"><p>PCR of insert fragment [B0034+Zif268+AlsS] NOT OK</p></li>
 
-
<li class="green e4"><p>transformation of B0034+Zif268+AlsS and cultivation on LB-A plate</p></li>
 
-
<li class="green e4"><p>mini-prep of cultivated B0034 and ilvD E. coli</p></li>
 
-
<li class="green e4"><p>Single colony isolation from HivC LB-A plate, and cultivation in liquid LB-A</p></li>
 
</ul>
</ul>
</div>
</div>
Line 738: Line 693:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>24</p></div>
+
<div class="daybar"><p>23</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 748: Line 701:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>mini-prep of cultivated HivC E. coli</p></li>
 
-
<li class="green e2"><p>digestion : B0034 [SP] &amp; ilvD [XP]</p></li>
 
</ul>
</ul>
</div>
</div>
Line 755: Line 706:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>25</p></div>
+
<div class="daybar"><p>24</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 766: Line 714:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>digestion : pSB1C3 [EP] &amp; HivC [ES] &amp; HivC [SP]</p></li>
 
-
<li class="green e3"><p>ligation :insert HivC [ES] &amp; ilvD [XP]/Vector pSB1C3 [EP]</p></li>
 
-
<li class="green e3"><p>ligation :insert ilvD [XP]/Vector HivC+pSB1A3 [SP]</p></li>
 
</ul>
</ul>
</div>
</div>
Line 774: Line 719:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>26</p></div>
+
<div class="daybar"><p>25</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 783: Line 727:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of HivC+ilvD and cultivation on LB-A &amp; LB-C plate</p></li>
 
</ul>
</ul>
</div>
</div>
Line 789: Line 732:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>27</p></div>
+
<div class="daybar"><p>26</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 798: Line 740:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>PCR of insert fragment [B0034+Zif268+AlsS] NOT OK</p></li>
 
</ul>
</ul>
</div>
</div>
Line 804: Line 745:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>28</p></div>
+
<div class="daybar"><p>27</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 815: Line 753:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>transformation of pSB1A3 &amp; pSB1C3 and cultivation on LB-A &amp; LB-C plate</p></li>
 
-
<li class="green e3"><p>PCR of insert fragment [HivC+ilvD] OK</p></li>
 
-
<li class="green e3"><p>DNA sequencing NOT OK</p></li>
 
</ul>
</ul>
</div>
</div>
Line 827: Line 762:
<div class="week">
<div class="week">
<div class="day">
<div class="day">
-
<div class="daybar"><p>29</p></div>
+
<div class="daybar"><p>28</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
Line 836: Line 771:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>Single colony isolation from pSB1C3 LB-C plate, and cultivation in liquid LB-C</p></li>
+
<li class="green"><p>Transformation of PompC mini and cultivation on LB-A plate</p></li>
</ul>
</ul>
</div>
</div>
Line 842: Line 777:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>30</p></div>
+
<div class="daybar"><p>29</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 851: Line 787:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>mini-prep of cultivated &amp; pSB1C3 E. coli</p></li>
+
<li class="green e2"><p>transformation of PompC , Ar mini and cultivation on LB-A  plate</p></li>
 +
<li class="green e2"><p>single colony isolation from PompC , Ar</p></li>
</ul>
</ul>
</div>
</div>
Line 857: Line 794:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p></p></div>
+
<div class="daybar"><p>30</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 865: Line 804:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e2"><p>Transformation of 37ﾟC RBS and ho1.</p></li>
 +
<li class="green e2"><p>Mini-prep of cultivated PompC , Ar  E.coli</p></li>
</ul>
</ul>
-
</div>
+
</div>
-
 
+
</div>
</div>
<div class="day">
<div class="day">
Line 945: Line 885:
<div class="daysmonth">
<div class="daysmonth">
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week mweek">
+
<div class="week bigwk">
 +
<div class="day">
 +
<div class="daybar"><p></p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
<div class="day">
<div class="day">
<div class="daybar"><p></p></div>
<div class="daybar"><p></p></div>
Line 976: Line 929:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 984: Line 938:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>digestion : pLac [EP] &amp; pSB1K3 [EP]</p></li>
+
<li class="green e4"><p>Cultivation of 37ﾟC RBS and ho1</p></li>
-
<li class="green e3"><p>ligation :insert pLac [EP]/Vector pSB1K3 [EP]</p></li>
+
<li class="green e4"><p>Digestion: [pSB1C3] EP&[B0030]ES&{pcyA]XP</p></li>
-
<li class="green e3"><p>transformation of pLac+pSB1K3 and cultivation on LB-K plate</p></li>
+
<li class="green e4"><p>Transformation of pSB1A3 &pSB1K3</p></li>
 +
<li class="green e4"><p>Transformation of mGFP[pSB1A2] and cultivation on LB-A plate.</p></li>
</ul>
</ul>
</div>
</div>
Line 995: Line 950:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,004: Line 963:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e4"><p>PCR of insert fragment [pLac+pSB1K3] OK</p></li>
+
<li class="green e8"><p>Cultivation of PompC&B0034&LacI&Plac&B0030&TetR&J61048&B0015 Ar E.coli in liquid LB-A tubes.</p></li>
-
<li class="green e4"><p>DNA Sequencing OK</p></li>
+
<li class="green e8"><p>Cultivation of pSB1A3&pSB1K3 with liquid LB</p></li>
-
<li class="green e4"><p>transformation of B0034_new and cultivation on LB-A plate</p></li>
+
<li class="green e8"><p>Mini-prep of  37ﾟC RBS and ho1──Fail</p></li>
-
<li class="green e4"><p>Single colony isolation from B0034 LB-A plate, and cultivation in liquid LB-A</p></li>
+
<li class="green e8"><p>Ligation: [pSB1C3] EP&[B0030]ES& {pcyA]XP</p></li>
 +
<li class="green e8"><p>Transformation of B0030</p></li>
 +
<li class="green e8"><p>Cultivation of 37ﾟC RBS and ho1 with liquid LB</p></li>
 +
<li class="green e8"><p>Transformation of RBS+pcyA+psB1C3.</p></li>
 +
<li class="green e8"><p>Transformation of mGFP[pSB1A2] and cultivation on LB-A plate</p></li>
</ul>
</ul>
</div>
</div>
Line 1,016: Line 979:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,026: Line 992:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e5"><p>mini-prep of cultivated B0034 E. coli</p></li>
+
<li class="green e8"><p>Mini-prep of cultivated PompC &B0034&LacI&Plac&B0030&TetR&J61048&B0015,and then digestion:[PompC ]ES&[B0034]XP&[LacI]ES&[Plac]XP&[B0030]ES&[TetR]ES&[TetR]XP&[J61048]XP&[J61048]ES&[B0015]XP.</p></li>
-
<li class="green e5"><p>digestion : B0034 [SP] </p></li>
+
<li class="green e8"><p>Mini-prep of ho1,pSB1A3, pSB1K3 and 37ﾟC RBS</p></li>
-
<li class="green e5"><p>ligation :insert Zif268+AlsS [XP]/Vector B0034+pSB1A3 [SP]</p></li>
+
<li class="green e8"><p>Digestion: [B0030+pcyA]XP</p></li>
-
<li class="green e5"><p>transformation of B0034+Zif268+AlsS</p></li>
+
<li class="green e8"><p>Cultivation of B0030 with liquid LB-C</p></li>
-
<li class="green e5"><p>cultivation on LB-A plate</p></li>
+
<li class="green e8"><p>Check RBS+pcyA+psB1C3(PCR & Electrophoresis)──FAIL</p></li>
 +
<li class="green e8"><p>Digestion: [Pcons]ES& [ho1]XP.</p></li>
 +
<li class="green e8"><p>Single colony isolation from 37。C RBS Ar,pSB1A3,pSB1C3 and pSB1K3</p></li>
 +
<li class="green e8"><p>electrophoresis of mGFP dig E</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
+
<div class="day brn">
<div class="daybar"><p>4</p></div>
<div class="daybar"><p>4</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,046: Line 1,021:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [B0034+Zif268+AlsS]-----self ligation</p></li>
+
<li class="green e8"><p>Electrophoresis of [PompC]ES&[B0030]ES&[LacI]ES&[TetR]ES&[J61048]XP&[TetR]ES&[B0015]XP OK</p></li>
-
<li class="green e2"><p>transformation of 37℃ RBS and cultivation on LB-C plate</p></li>
+
<li class="green e8"><p>Ligation: insert [PompC]ES+[B0030]XP&[LacI]ES+[J61048]XP&[J61048]ES+[Plac]XP&[B0030]ES+[TetR]XP&[TetR]ES+[B0015]XP/vector[PSB1C3]EP</p></li>
 +
<li class="green e8"><p>Transformation of B0030+ho1.</p></li>
 +
<li class="green e8"><p>Mini-prep of cultivated 37。C RBS Ar pSB1A3,37。C RBS+mGFP, Kr  and pSB1K3 E.coli</p></li>
 +
<li class="green e8"><p>digestion:37。C RBS dig ES,mGFP dig XP,LuxR dig XP and pSB1K3 dig EP</p></li>
 +
<li class="green e8"><p>electrophoresis of digestion products of LuxR,37。C RBS, mGFP and pSB1K3</p></li>
 +
<li class="green e8"><p>ligation:37。C RBS+luxR+pSB1K3</p></li>
 +
<li class="green e8"><p>transformation of 37。C RBS+mGFP, Kr->pSB1K3 and cultivation on LB-K plate</p></li>
</ul>
</ul>
</div>
</div>
-
</div>
+
</div>
-
<div class="day brn">
+
</div>
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week bigwk">
 +
<div class="day">
<div class="daybar"><p>5</p></div>
<div class="daybar"><p>5</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,061: Line 1,052:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e7"><p>Electrophoresis of digested RBS+pcyA──OK</p></li>
 +
<li class="green e7"><p>Ligation: RBS+ho1 </p></li>
 +
<li class="green e7"><p>Cultivation of J61048 with liquid LB-C</p></li>
 +
<li class="green e7"><p>Digestion: [pSB1A3]EP &[pSB1K3]EP</p></li>
 +
<li class="green e7"><p>Transformation of ligated RBS+ho1.</p></li>
 +
<li class="green e7"><p>Ligation:37。C RBS+luxR+pSB1K3 and 37。C RBS+mGFP+pSB1K3</p></li>
 +
<li class="green e7"><p>transformation of  37。C RBS+luxR, Kr and 37。C RBS+mGFP ,Kr and cultivation on LB-K plate</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
</div>
 
-
<!---------------------------------------- week start ---------------------------------------->
 
-
<div class="week mweek">
 
<div class="day">
<div class="day">
<div class="daybar"><p>6</p></div>
<div class="daybar"><p>6</p></div>
<div class="dots">
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
</ul>
</div>
</div>
<div class="open">
<div class="open">
 +
<ul>
 +
<li class="green e6"><p>Mini-prep of J61048</p></li>
 +
<li class="green e6"><p>Ligation: RBS+pcyA+Pcons&pSB1K3</p></li>
 +
<li class="green e6"><p>Electrophoresis of J61048.</p></li>
 +
<li class="green e6"><p>Digestion: [J61048]XP</p></li>
 +
<li class="green e6"><p>Transformation of RBS+pcyA+Pcons.</p></li>
 +
<li class="green e6"><p>Check LB-A&C&K plates.</p></li>
 +
</ul>
</div>
</div>
Line 1,081: Line 1,092:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,086: Line 1,101:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>Mini-prep of J61048
 +
-Ligation: RBS+pcyA+Pcons&pSB1K3</p></li>
 +
<li class="green e4"><p>Electrophoresis of J61048.</p></li>
 +
<li class="green e4"><p>Digestion: [J61048]XP</p></li>
 +
<li class="green e4"><p>Transformation of RBS+pcyA+Pcons.-Check LB-A&C&K plates.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,094: Line 1,114:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,100: Line 1,122:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of 37℃ RBS and cultivation on LB-Kplate</p></li>
+
<li class="green e3"><p>Ligation:insert[PompC]ES+[B0034]XP&[LacI]ES+[J61048]XP&[B0030]ES+[TetR]XP&[TetR]ES+[B0015]XP/vector[PSB1C3]E</p></li>
 +
<li class="green e3"><p>PCR of 37。RBS+luxR, Kr and 37。C RBS+mGFP ,Kr</p></li>
 +
<li class="green e3"><p>electrophoresis of 37。RBS+luxR ,Kr and 37。C RBS+mGFP ,Kr</p></li>
</ul>
</ul>
</div>
</div>
Line 1,106: Line 1,130:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>9</p></div>
+
<div class="daybar"><p>9</p></div> <div class="dots">
-
<div class="dots">
+
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,115: Line 1,141:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>Single colony isolation from 37℃ RBS LB-K plate, and cultivation in liquid LB-K</p></li>
+
<li class="green e4"><p>Transformation and cultivation of PompC+B0034&LacI+J61048&B0030+TetR&TetR+B0015(PSB1C3) on LB-C plates.</p></li>
 +
<li class="green e4"><p>PCR of 37。RBS+luxR, Kr and 37。C RBS+mGFP ,Kr</p></li>
 +
<li class="green e4"><p>electrophoresis of 37。RBS+luxR, Kr and 37。C RBS+mGFP ,Kr</p></li>
 +
<li class="green e4"><p>single colony isolation from37。RBS+luxR, Kr and 37。C RBS+mGFP ,Kr</p></li>
</ul>
</ul>
</div>
</div>
Line 1,124: Line 1,153:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,131: Line 1,161:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>digestion : B0034 [ES] &amp; pSB1C3 [EP]</p></li>
+
<li class="green e3"><p>Cultivation of Pcons+RBS+pcyA and B0030 with liquid LB.</p></li>
-
<li class="green e2"><p>mini-prep of cultivated 37℃ RBS E. coli</p></li>
+
<li class="green e3"><p>Mini-prep of cultivated 37。C RBS+luxR, Kr E.coli and 37。C RBS+mGFP ,Kr E.coli</p></li>
 +
<li class="green e3"><p>digestion: 37。RBS+luxR, dig ES and 37。C RBS+mGFP dig ES</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
+
<div class="day brn">
<div class="daybar"><p>11</p></div>
<div class="daybar"><p>11</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,148: Line 1,180:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>ligation :insert B0034 [ES] &amp; pSB1C3 [EP]/Vector pSB1C3 [EP]</p></li>
+
<li class="green e3"><p>Mini-prep of Pcons+RBS+pcyA.</p></li>
-
<li class="green e2"><p>transformation of B0034+Zif268+AlsS and cultivation on LB-C plate</p></li>
+
<li class="green e3"><p>Electrophoresis of Pcons+RBS+pcyA──NOT OK.</p></li>
 +
<li class="green e3"><p>Electrophoresis of 37。C RBS+mGFP, Kr mini, 37。C RBS+mGFP dig ES, 37。C RBS+luxR, Kr mini and 37。C RBS+luxR dig ES</p></li>
</ul>
</ul>
</div>
</div>
-
</div>
+
</div>
-
<div class="day brn">
+
</div>
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week mweek">
 +
<div class="day">
<div class="daybar"><p>12</p></div>
<div class="daybar"><p>12</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,167: Line 1,200:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e4"><p>digestion : pLac [ES] &amp; pLac [SP]</p></li>
+
<li class="green"><p>Electrophoresis of 37。C+luxR mini, dig ES and 37。C RBS+mGFP mini,dig ES</p></li>
-
<li class="green e4"><p>digestion : B0034 [SP] Kr (gel extraction) , B0034 [SP] Ar (gel extraction) , 37℃ RBS [XP] &amp; HivC [XP]</p></li>
+
-
<li class="green e4"><p>ligation : (1. 2 parts) insert HivC [XP], Vector B0034+pSB1K3 [SP]</p></li>
+
-
<li class="green e4"><p>ligation : (2. 3 parts) insert HivC [XP], Vector B0034+pSB1A3 [SP]</p></li>
+
</ul>
</ul>
</div>
</div>
</div>
</div>
-
</div>
 
-
<!---------------------------------------- week start ---------------------------------------->
 
-
<div class="week bigwk">
 
<div class="day">
<div class="day">
<div class="daybar"><p>13</p></div>
<div class="daybar"><p>13</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,187: Line 1,219:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e5"><p>Cultivation of TetR+B0015 Cr E.coli in liquid LB-C tube</p></li>
 +
<li class="green e5"><p>Single colony isolation of backbone PSB1K3 E.coli</p></li>
 +
<li class="green e5"><p>Ligation:insert[B0030]ES+[TetR]XP&[TetR]ES+[B0015]XP/vector[PSB1C3]EP.</p></li>
 +
<li class="green e5"><p>Electrophoresis of Pcons+RBS+pcyA──NOT OK</p></li>
 +
<li class="green e5"><p>Transformation of B0032&B0034.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,195: Line 1,232:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,200: Line 1,239:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e2"><p>Ligation:insert[PompC]ES+[B0034]XP&[LacI]ES+[J61048]XP/vector[PSB1C3]EP. </p></li>
 +
<li class="green e2"><p>Cultivation of Pcons+RBS+pcyA & B0032&B0034 with liquid LB.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,208: Line 1,249:
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,216: Line 1,256:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>PCR of insert fragment [B0034+Zif268+AlsS] OK</p></li>
+
<li class="green e2"><p>Mini-prep of cultivated PSB1K3 E.coli </p></li>
-
<li class="green e3"><p>DNA Sequencing OK</p></li>
+
<li class="green e2"><p>Cultivation of B0015+TetR+PSB1C3 and TetR+B0015+PSB1C3 in liquid LB-C plates and Plac+PSB1A3 in liquid LB-A plate. Mini-prep of Pcons+RBS+pcyA&B0032&B0034.</p></li>
-
<li class="green e3"><p>transformation of B0034+ HivC and cultivation on LB-A &amp; LB-K plate</p></li>
+
</ul>
</ul>
</div>
</div>
Line 1,227: Line 1,266:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,232: Line 1,273:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e2"><p>Ligation:insert[PompC]ES+[B0034]XP&[LacI]ES+[J61048]XP/vector[PSB1K3]EP</p></li>
 +
<li class="green e2"><p>Transformation and cultivation of PompC+B0034&LacI+J61048 (PSB1K3) on LB-K plates.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,240: Line 1,283:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,245: Line 1,292:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>cultivation of PompC+B0034&LacI+J61048 Kr E.coli on liquid LB-K tubes.</p></li>
 +
<li class="green e4"><p>single colony isolation of PompC+B0034&LacI+J61048 Kr E.coli.</p></li>
 +
<li class="green e4"><p>Electrophoresis of Pcons+RBS+pcyA(NOT OK) &B0032&B0034</p></li>
 +
<li class="green e4"><p>Cultivation of Pcons+RBS+pcyA.Digestion: [B0032]ES &[B0023]ES.</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
+
<div class="day brn">
<div class="daybar"><p>18</p></div>
<div class="daybar"><p>18</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,261: Line 1,313:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>Single colony isolation from pSB1A3 LB-A plate, and cultivation in liquid LB-A</p></li>
+
<li class="green e4"><p>Cultivation of PompC+B0034&LacI+J61048 Kr E.coli from single colony isolation plate made yesterday</p></li>
-
<li class="green e3"><p>Single colony isolation from B0034+Zif268+AlsS LB-C plate, and cultivation in liquid LB-C</p></li>
+
<li class="green e4"><p>Min-prep of  cultivated [B0030+TetR]Cr&[ Plac]Ar&[PompC+B0034]Kr&[LacI+J61048]Kr  E.coli and then digestion:[PompC+B0034]ES &[LacI+J61048]XP. Mini-prep Pcons+RBS+pcyA</p></li>
-
<li class="green e3"><p>Single colony isolation from B0034+HivC LB-A &amp; K plate, and cultivation in liquid LB-A &amp; K</p></li>
+
<li class="green e4"><p>Electrophoresis of Pcons+RBS+pcyA& digested B0032&B0034──NOT OK</p></li>
 +
<li class="green e4"><p>Digestion: [B0032]ES &[B0034]ES &[Pcons+RBS+pcyA]E.</p></li>
</ul>
</ul>
</div>
</div>
-
</div>
+
</div>
-
<div class="day brn">
+
</div>
 +
<!---------------------------------------- week end ---------------------------------------->
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week bigwk">
 +
<div class="day">
<div class="daybar"><p>19</p></div>
<div class="daybar"><p>19</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,283: Line 1,337:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e6"><p>mini-prep of cultivated pSB1A3 E. coli &amp; B0034+Zif268+AlsS</p></li>
+
<li class="green e3"><p>Electrophoresis of [LacI+J61048]mini & dig XP and [PompC+B0034]mini & dig ES and [PSB1K3]mini & dig EP.</p></li>
-
<li class="green e6"><p>digestion: pSB1A3 [EP] &amp; B0034+Zif268+AlsS [XP] </p></li>
+
<li class="green e3"><p>Electrophoresis of digested B0032&B0034&Pcons+RBS+pcyA──OK</p></li>
-
<li class="green e6"><p>ligation: (1.3 parts)insert pLac [ES] &amp; B0034+Zif268+AlsS [XP], Vector pSB1A3 [EP]</p></li>
+
<li class="green e3"><p>Transformation of pSB1C3</p></li>
-
<li class="green e6"><p>ligation: (2.2 parts)insert B0034+Zif268+AlsS [XP], Vector pLac+pSB1K3 [SP]</p></li>
+
-
<li class="green e6"><p>transformation of pLac+B0034+Zif268+AlsS and cultivation on LB-A plate &amp; LB-K plate</p></li>
+
-
<li class="green e6"><p>mini-prep of cultivated B0034+HivC (Ar &amp; Kr)</p></li>
+
</ul>
</ul>
</div>
</div>
</div>
</div>
-
</div>
 
-
<!---------------------------------------- week end ---------------------------------------->
 
-
<!---------------------------------------- week start ---------------------------------------->
 
-
<div class="week">
 
<div class="day">
<div class="day">
<div class="daybar"><p>20</p></div>
<div class="daybar"><p>20</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,308: Line 1,357:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [pLac+B0034+Zif268+AlsS] OK</p></li>
+
<li class="green e4"><p>Digestion:[Plac]P.</p></li>
-
<li class="green e2"><p>DNA Sequencing 3 parts OK</p></li>
+
<li class="green e4"><p>Ligation: Pcons+RBS+pcyA&pSB1K3, B0032+ho1&pSB1C3, B0034+ho1&pSB1C1 </p></li>
 +
<li class="green e4"><p>Transformation of Pcons+RBS+pcyA&pSB1K3, B0032+ho1&pSB1C3, B0034+ho1&pSB1C1</p></li>
 +
<li class="green e4"><p>Single colony isolation from pSB1C3</p></li>
</ul>
</ul>
</div>
</div>
Line 1,318: Line 1,369:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,324: Line 1,378:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>digestion : 37℃ RBS [XP] , ilvD [ES] &amp; pSB1C3 [EP]</p></li>
+
<li class="green e4"><p>Single colony isolation of TetR+B0015 Cr Ecoli</p></li>
 +
<li class="green e4"><p>Electrophoresis of [PompC+B0034]mini&dig ES and[Plac]mini&dig P.</p></li>
 +
<li class="green e4"><p>Cultivation of Pcons+RBS+pcyA &B0032+ho1 with liquid LB</p></li>
 +
<li class="green e4"><p>Mini-prep of cultivated pSB1C3 E.coli</p></li>
</ul>
</ul>
</div>
</div>
Line 1,333: Line 1,390:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,338: Line 1,402:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e7"><p>Digestion:[Plac]XP</p></li>
 +
<li class="green e7"><p>Electrophoresis of [PompC+B0034]mini&dig ES and[Plac]mini&dig XP</p></li>
 +
<li class="green e7"><p>Ligation: insert[TetR]ES+[B]XP/vector[PSBC3]EP</p></li>
 +
<li class="green e7"><p>Mini-prep of Pcons+RBS+pcyA</p></li>
 +
<li class="green e7"><p>Mini-prep of cultivated 37。RBS+mGFP E.coli</p></li>
 +
<li class="green e7"><p>digestion:37。C RBS+mGFP dig ES and mini</p></li>
 +
<li class="green e7"><p>electrophoresis of 37。C RBS+mGFP mini and 37。C RBS+mGFP dig ES.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,353: Line 1,424:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>digestion : HivC+ilvD-3,18 [EP]</p></li>
+
<li class="green e2"><p>Single colony isolation of TetR+B0015  transformation of TetR +B0015 +PSB1C3 on LB-C plates</p></li>
-
<li class="green e2"><p>ligation :insert ilvD[ES] &amp; 37℃ RBS [XP], Vector pSB1C3 [EP]</p></li>
+
<li class="green e2"><p>Ligation +transformation on: insert[TetR]ES+[B0015]XP&[PompC]ES+[B0034]/vector[PSB1K3]EP on LB-K plates</p></li>
</ul>
</ul>
</div>
</div>
Line 1,363: Line 1,434:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,369: Line 1,444:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>Single colony isolation from pLac+B0034+Zif268+AlsS LB-A plate, and cultivation in liquid LB-A</p></li>
+
<li class="green e5"><p>Single colony isolation of PompC+B0034+PSB1K3 E.coli</p></li>
 +
<li class="green e5"><p>Electrophoresis of Pcons+RBS+pcyA.</p></li>
 +
<li class="green e5"><p>-Ligation: B0032+ho1&B0034+ho1.</p></li>
 +
<li class="green e5"><p>Digestion; [Pcons+RBS+pcyA]ES &[ho1]XP&[pSB1C3]EP.</p></li>
 +
<li class="green e5"><p>Electrophoresis of 37。 RBS+mGFP dig ES</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
+
<div class="day brn">
<div class="daybar"><p>25</p></div>
<div class="daybar"><p>25</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,385: Line 1,470:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>mini-prep of cultivated pLac+B0034+Zif268+AlsS E. coli</p></li>
+
<li class="green e8"><p>Ligation and transformation: insert [PompC]ES+[B0034]XP/vector[PSB1K3]EP on LB-K plates</p></li>
-
<li class="green e2"><p>transformation of ilvD+37℃ RBS and cultivation on LB-C plate</p></li>
+
<li class="green e8"><p>Single colony isolation of PompC+B0034+PSB1K3 E.coli again</p></li>
 +
<li class="green e8"><p>Mini-prep of cultivated TetR+B0015 Cr  E.coli and digestion:[TetR+B0015]P</p></li>
 +
<li class="green e8"><p>Electrophoresis of :[TetR+B0015]P</p></li>
 +
<li class="green e8"><p>Digestion: [TetR+B0015]XP</p></li>
 +
<li class="green e8"><p>Cultivation of PompC+B0034 in liquid LB-K tubes.</p></li>
 +
<li class="green e8"><p>Mini-prep of B0032+ho1</p></li>
 +
<li class="green e8"><p>Cultivation of B0034+ho1 with liquid LB-A</p></li>
</ul>
</ul>
</div>
</div>
-
</div>
+
</div>
-
<div class="day brn">
+
</div>
 +
<!---------------------------------------- week end ---------------------------------------->
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week bigwk">
 +
<div class="day">
<div class="daybar"><p>26</p></div>
<div class="daybar"><p>26</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,402: Line 1,499:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [ilvD+37℃]-----OK</p></li>
+
<li class="green e4"><p>Cultivation of PompC+B0034Kr E.coli  in liquid LB-K tubes again</p></li>
-
<li class="green e2"><p>DNA Sequencing OK</p></li>
+
<li class="green e4"><p>Mini-prep of cultivate PompC+B0034 Kr E.coli</p></li>
 +
<li class="green e4"><p>Electrophoresis of [TetR+B0015] dig XP not OK</p></li>
 +
<li class="green e4"><p>Mini-prep of B0034+ho1
 +
Electrophoresis of  Pcons+RBS+pcyA & B0032+ho1 & B0034+ho1.</p></li>
</ul>
</ul>
</div>
</div>
-
</div>
+
</div>
-
</div>
+
-
<!---------------------------------------- week end ---------------------------------------->
+
-
<!---------------------------------------- week start ---------------------------------------->
+
-
<div class="week mweek">
+
<div class="day">
<div class="day">
<div class="daybar"><p>27</p></div>
<div class="daybar"><p>27</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,421: Line 1,523:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e6"><p>Digestion:[TetR+B0015]XP</p></li>
 +
<li class="green e6"><p>Electrophoresis of [TetR+B0015]mini&dig XP</p></li>
 +
<li class="green e6"><p>Digestion:[PompC+B0034]E first</p></li>
 +
<li class="green e6"><p>Electrophoresis of [PompC+B0034]E</p></li>
 +
<li class="green e6"><p>Digestion: [PompC+B0034]E, S later</p></li>
 +
<li class="green e6"><p>Ligation: insert[PompC]ES+[B0034]XP/vector[PSB1K3]EP</p></li>
</ul>
</ul>
</div>
</div>
Line 1,429: Line 1,537:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,434: Line 1,546:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>Electrophoresis of [PompC+B0034]ES not OK</p></li>
 +
<li class="green e4"><p>Transformation of PompC+B0034+PSB1K3.</p></li>
 +
<li class="green e4"><p>Electrophoresis of B0034+ho1</p></li>
 +
<li class="green e4"><p>Digestion: [B0034+ho1]XP &[Pcons+RBS+pcyA]E</p></li>
</ul>
</ul>
</div>
</div>
Line 1,442: Line 1,558:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,447: Line 1,564:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green"><p>PCR and electrophoresis test. Electrophoresis of digested B0034+ho1 &Pcons+RBS+pcyA.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,455: Line 1,573:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,462: Line 1,584:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Single colony isolation from PBSII+ilvC LB-K plate, and cultivation in liquid LB-K</p></li>
+
<li class="green e6"><p>Cultivation of PompC+B0034+PSB1K3 E.coli in liquid LB-K tubes</p></li>
-
<li class="green e2"><p>ligation : insert HivC [XP], vector B0034+pSB1A3 [SP]</p></li>
+
<li class="green e6"><p>Ligation: insert[TetR]ES+[B0015]XP/vector[PSB1K3]EP</p></li>
 +
<li class="green e6"><p>Transformation of  Plac+PSB1A3 on LB-A plate</p></li>
 +
<li class="green e6"><p>Digestion:B0015 Ar dig XP, pSB1A3 Ar dig EP and pSB1C3 Cr dig EP</p></li>
 +
<li class="green e6"><p>ligation:37。C RBS+mGFP+ter. [B0015], Cr</p></li>
 +
<li class="green e6"><p>transformation of 37。C RBS+mGFP+ter., Cr</p></li>
</ul>
</ul>
</div>
</div>
Line 1,472: Line 1,598:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,482: Line 1,610:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e5"><p>mini-prep of cultivated PBSII+ilvC E. coli</p></li>
+
<li class="green e7"><p>Mini-prep of cultivated PompC+B0034 Kr E.coli, then</p></li>
-
<li class="green e5"><p>digestion : pSB1C3 [EP] &amp; PBSII+ilvC [XP] </p></li>
+
<li class="green e7"><p>Digestion  and electrophoresis of [PompC+B0034]E</p></li>
-
<li class="green e5"><p>ligation : insert B0034 [ES] &amp; PBSII+ilvC [XP], vector pSB1C3 [EP]</p></li>
+
<li class="green e7"><p>Decide to determine the sequence of PompC+B0015.</p></li>
-
<li class="green e5"><p>transformation of B0034+PBSII+ilvC and cultivation on LB-C plate</p></li>
+
<li class="green e7"><p>Cultivation of ho1 with liquid LB-K and pSB1C3 with liquid LB-C.</p></li>
-
<li class="green e5"><p>transformation of B0034+HivC and cultivation on LB-A plate</p></li>
+
<li class="green e7"><p>PCR of 37。C RBS+mGFP+ter. Cr</p></li>
 +
<li class="green e7"><p>electrophoresis of 37。C RBS+mGFP+ter., ter.[B0015] digXP, pSB1C3 dig EP and pSB1A3 dig EP</p></li>
 +
<li class="green e7"><p>transformation of BBa _E1010 Kr</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
 
-
<div class="daybar"><p></p></div>
 
-
<div class="dots">
 
-
<ul>
 
-
</ul>
 
-
</div>
 
-
 
-
<div class="open">
 
-
<ul>
 
-
</ul>
 
-
</div>
 
-
 
-
</div>
 
<div class="day brn">
<div class="day brn">
<div class="daybar"><p></p></div>
<div class="daybar"><p></p></div>
Line 1,516: Line 1,633:
</div>
</div>
-
</div>
+
</div>
</div>
</div>
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week end ---------------------------------------->
Line 1,541: Line 1,658:
<div class="daysmonth">
<div class="daysmonth">
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week">
+
<div class="week mweek">
<div class="day">
<div class="day">
<div class="daybar"><p></p></div>
<div class="daybar"><p></p></div>
Line 1,608: Line 1,725:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>1</p></div>
+
<div class="daybar"><p></p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 1,618: Line 1,733:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [B0034+PBSII+ilvC] OK!</p></li>
 
-
<li class="green e2"><p>DNA Sequencing OK!</p></li>
 
</ul>
</ul>
</div>
</div>
Line 1,625: Line 1,738:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>2</p></div>
+
<div class="daybar"><p>1</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,633: Line 1,752:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e6"><p>PCR and single colony isolation of TetR+B0015+PSB1K3</p></li>
 +
<li class="green e6"><p>Electrophoresis of PCR product made at this morning and Plac(5/30 Ar) OK</p></li>
 +
<li class="green e6"><p>Digestion:[Ter(B0015)]XP</p></li>
 +
<li class="green e6"><p>Transformation of Ter(B0015) on LB-A plate & mGFP on LB-K plate & 37oC RBS+mGFP+Ter(B0015) on LB-C plate</p></li>
 +
<li class="green e6"><p>Ligation: insert 37oC RBS[ES] & Ter(B0015)[XP]/ vector pSB1C3[EP]</p></li>
 +
<li class="green e6"><p>Single colony isolation from Ter(B0015) LB-A plate, and cultivation in liquid LB-A</p></li>
</ul>
</ul>
</div>
</div>
Line 1,639: Line 1,764:
</div>
</div>
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week mweek">
+
<div class="week bigwk">
<div class="day">
<div class="day">
-
<div class="daybar"><p>3</p></div>
+
<div class="daybar"><p>2</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,650: Line 1,779:
<div class="open">
<div class="open">
-
                                                                <ul>
+
<ul>
-
<li class="green e2"><p>mini-prep of cultivated B0034+PBSII+ilvC E. coli &amp; pLac+B0034+Zif268+AlsS E. coli</p></li>
+
<li class="green e6"><p>Ligation: insert [TetR]ES+[B0015]XP/vector [PSB1K3]EP and transformation of this part.</p></li>
-
                                                                </ul>
+
<li class="green e6"><p>Ligation: B0032+ho1&B0034+ho1</p></li>
-
                                                                <ul>
+
<li class="green e6"><p>Transformation of B0032+ho1&B0034+ho1</p></li>
-
<li class="green e2"><p>mini-prep of cultivated HivC+ilvD-12,20 E. coli </p></li>
+
<li class="green e6"><p>Cultivation of pSB1C3(C20&C10)with liquid LB.</p></li>
-
                                                                </ul>
+
<li class="green e6"><p>PCR of insert fragment[37oC RBS+mGFP+Ter(B0015)]</p></li>
 +
<li class="green e6"><p>Transformation of mRFP on LB-A plate& LB-C plate& LB-K plate-Mini-prep of cultivated Ter(B0015) E.coli.</p></li>
 +
</ul>
</div>
</div>
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>4</p></div>
+
<div class="daybar"><p>3</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,669: Line 1,806:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>digestion : HivC+ilvD-12,20 [XP]</p></li>
+
<li class="green e7"><p>PCR of Plac &TetR+B0015</p></li>
 +
<li class="green e7"><p>Mini-prep of pSB1C3</p></li>
 +
<li class="green e7"><p>Cultivation of B0032+ho1&B0034+ho1 with LB-A plate PCR of B0032+ho1&B0034+ho1</p></li>
 +
<li class="green e7"><p>Electrophoresis of B0032+ho1&B0034+ho1──NOT OK.</p></li>
 +
<li class="green e7"><p>Single colony isolation from 37oC RBS+mGFP+Ter(B0015) LB-K plate, and cultivation in liquid LB-K & mRFP LB-A, and cultivation in liquid LB-K</p></li>
 +
<li class="green e7"><p>Mini-prep of cultivated 37oC RBS+mGFP+Ter(B0015) E.coli</p></li>
 +
<li class="green e7"><p>Digestion:[37oC RBS+mGFP+Ter(B0015)]XP</p></li>
</ul>
</ul>
</div>
</div>
Line 1,675: Line 1,818:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>5</p></div>
+
<div class="daybar"><p>4</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,684: Line 1,834:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>Design the primer for pLac+B0034+Zif268+AlsS point mutation (pm)</p></li>
+
<li class="green e8"><p>Electrophoresis of  Plac .TetR+B0015 OK</p></li>
 +
<li class="green e8"><p>Cultivation of Plac.TetR+B0015 in liquid LB-A tubes. PCR of B0032+ho1&B0034+ho1</p></li>
 +
<li class="green e8"><p>Electrophoresis of B0032+ho1(OK)&B0034+ho1(After PCR)</p></li>
 +
<li class="green e8"><p>Mini-prep of B0032+ho1. Digestion: [B0032+ho1]XP.</p></li>
 +
<li class="green e8"><p>Cultivation of ho1 with liquid LB-K</p></li>
 +
<li class="green e8"><p>Single colony isolation from mRFP LB-A plate, and cultivation in liquid LB-A</p></li>
 +
<li class="green e8"><p>Digestion:[37oC RBS+mGFP+Ter(B0015)]XP</p></li>
 +
<li class="green e8"><p>PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)].</p></li>
</ul>
</ul>
</div>
</div>
Line 1,690: Line 1,847:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>6</p></div>
+
<div class="daybar"><p>5</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,702: Line 1,863:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e4"><p>digestion : B0034 [ES] &amp; HivC [XP] &amp; pSB1C3 [EP] &amp; HivC [EP]</p></li>
+
<li class="green e8"><p>Mini-prep of cultivated Plac and TetR+B0015 E.coli and then conduct digestion :[Plac]P&[TetR+B0015]P</p></li>
-
<li class="green e4"><p>ligation : insert B0034 [ES] &amp; HivC [XP]/vector pSB1C3 [EP]</p></li>
+
<li class="green e8"><p>Electrophoresis of [Plac ]P and [TetR+B0015]P,[Plac]XP and [TetR+B0015]XP</p></li>
-
<li class="green e4"><p>ligation : insert HivC [EP]/vector pSB1C3 [EP]</p></li>
+
<li class="green e8"><p>Ligation: insert[PompC+B0034]ES+[LacI+J61048]XP/vector [PSB1A3]EP</p></li>
-
<li class="green e4"><p>transformation of B0034+ HivC &amp; HivC ,and cultivation on LB-C plate</p></li>
+
<li class="green e8"><p>Electrophoresis of B0032+ho1 mini&B0032+ho1(digested)&pSB1C3 mini&pSB1C3(digested)&ho1 mini&ho1(digested)</p></li>
 +
<li class="green e8"><p>Mini-prep of ho1</p></li>
 +
<li class="green e8"><p>Digestion: [ho1]XP</p></li>
 +
<li class="green e8"><p>Digestion:[37oC RBS+mGFP+Ter(B0015)]XP</p></li>
 +
<li class="green e8"><p>Transformation of mRFP on LB-A plate</p></li>
</ul>
</ul>
</div>
</div>
Line 1,711: Line 1,876:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>7</p></div>
+
<div class="daybar"><p>6</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,720: Line 1,891:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>PCR of insert fragment [B0034+ HivC] &amp; [HivC] OK</p></li>
+
<li class="green e7"><p>Transformation of PompC+B0034+LacI+J61048+PSB1A3</p></li>
 +
<li class="green e7"><p>Ligation: insert [PompC+B0034]ES+[LacI+J61048]XP/vector [PSB1A3]EP</p></li>
 +
<li class="green e7"><p>Electrophoresis of B0032+ho1 mini&B0032+ho1(digested)&ho1 mini&ho1 (digested).</p></li>
 +
<li class="green e7"><p>Single colony isolation from mRFP LB-A plate, and cultivation in liquid LB-A & LB-K & LB</p></li>
 +
<li class="green e7"><p>Digestion:[Ter(B0015)]EX</p></li>
 +
<li class="green e7"><p>Ligation: insert 37oC RBS+mGFP[ES]&Ter(B0015)[EX]</p></li>
 +
<li class="green e7"><p>Transformation of 37oCRBS+mGFP+Ter(B0015) on LB-K plate & LB-A plate, mRFP on LB-A plate</p></li>
</ul>
</ul>
</div>
</div>
Line 1,726: Line 1,903:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>8</p></div>
+
<div class="daybar"><p>7</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,736: Line 1,915:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Design the primer for B0034+PBSII+ilvC point mutation (pm)</p></li>
+
<li class="green e4"><p>Transformation of PompC+B0034+LacI+J61048 again</p></li>
-
<li class="green e2"><p>Single colony isolation from HivC, B0034+HivC &amp; ilvD+37℃ RBS LB-C plate, and cultivation in liquid LB-C</p></li>
+
<li class="green e4"><p>Single colony isolation of TetR+B0015 Kr E.coli</p></li>
 +
<li class="green e4"><p>Digestion:[LacI+J61048]ES. PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)]</p></li>
 +
<li class="green e4"><p>Single colony isolation from 37oCRBS+mGFP+Ter(B0015) LB-A plate, and cultivation in liquid LB-A.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,743: Line 1,924:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>9</p></div>
+
<div class="daybar"><p>8</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,754: Line 1,940:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>mini-prep of cultivated HivC &amp; B0034+HivC &amp; ilvD+37℃ RBS E. coli </p></li>
+
<li class="green e8"><p>PCR + single colony isolation of PompC+B0034+LacI+J61048 E.coli</p></li>
-
<li class="green e3"><p>Testing the temperature of the point mutation between of HivC &amp; ilvD by the m.p 50℃ of PCR</p></li>
+
<li class="green e8"><p>Electrophoresis of PompC+B0034+LacI+J61048 OK</p></li>
-
<li class="green e3"><p>ligation: insert B0034+HivC [ES] &amp; ilvD [XP]/vector pSB1A3 [EP];insert B0034+HivC [ES] &amp; ilvD+37℃ RBS [XP]/vector pSB1A3 [EP]
+
<li class="green e8"><p>Ligation: insert [LacI+J61048]ES+[Plac]XP/vector[PSB1C3]EP</p></li>
-
</p></li>
+
<li class="green e8"><p>Transformation of LacI+J61048+Plac on LB-C plate</p></li>
 +
<li class="green e8"><p>Cultivation of PompC+B0034+LacI+J61048 in LB-A tubes & TetR+B0015 in LB-K tube & PSB1A3 in LB-A tube &PSB1C3 in LB-C tube.</p></li>
 +
<li class="green e8"><p>Mini-prep of cultivated 37oCRBS+mGFP+Ter(B0015) E.coli</p></li>
 +
<li class="green e8"><p>Digestion:[ 37oCRBS+mGFP+Ter(B0015)]XP</p></li>
 +
<li class="green e8"><p>PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)].</p></li>
</ul>
</ul>
</div>
</div>
Line 1,764: Line 1,954:
</div>
</div>
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week mweek">
+
<div class="week bigwk">
<div class="day">
<div class="day">
-
<div class="daybar"><p>10</p></div>
+
<div class="daybar"><p>9</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,776: Line 1,968:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>transformation of B0034+ HivC+ ilvD &amp; B0034+ HivC+ ilvD+37℃ RBS ,and cultivation on LB-A plate</p></li>
+
<li class="green e4"><p>Mini-prep from cultivated PompC+B0034+LacI+J61048 Eco.li&PSB1C3 E.coli</p></li>
-
<li class="green e2"><p>Testing the temperature of the point mutation between of pLac+B0034+Zif268 &amp; AlsS by the m.p 55℃ of PCR failed</p></li>
+
<li class="green e4"><p>digestion : [PompC+B0034+LacI+J61048]ES&[PSB1C3]EP</p></li>
 +
<li class="green e4"><p>Electrophoresis of [PompC+B0034+LacI+J61048] ES& [LacI+J61048]ES&[PSB1C3]EP.</p></li>
 +
<li class="green e4"><p>Single colony isolation from 37oCRBS + mGFP+Ter(B0015)</p></li>
</ul>
</ul>
</div>
</div>
Line 1,783: Line 1,977:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>11</p></div>
+
<div class="daybar"><p>10</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,793: Line 1,988:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>DNA Sequencing B0034+HivC &amp; HivC OK</p></li>
+
<li class="green e3"><p>Mini-prep of cultivated 37oCRBS+mGFP+Ter(B0015)  E.coli</p></li>
-
<li class="green e2"><p>PCR of insert fragment [B0034+ HivC+ilvD] &amp; [B0034+ HivC+ilvD+37℃ RBS] OK</p></li>
+
<li class="green e3"><p>Digestion:[ 37oCRBS+mGFP+Ter(B0015)]XP</p></li>
 +
<li class="green e3"><p>Single colony isolation from 37oCRBS+mGFP+Ter(B0015) LB-A plate, and cultivation in liquid LB-A.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,800: Line 1,996:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>12</p></div>
+
<div class="daybar"><p>11</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
Line 1,810: Line 2,006:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Single colony isolation from B0034+ HivC+ilvD+37℃ RBS &amp; B0034+ HivC+ ilvD LB-A plate, and cultivation in liquid LB-A</p></li>
+
<li class="green e2"><p>PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)]</p></li>
-
<li class="green e2"><p>Single colony isolation from HivC(TA) LB-A plate, and cultivation in liquid LB-A</p></li>
+
<li class="green e2"><p>Transformation of mRFP on LB-K plate.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,817: Line 2,013:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>13</p></div>
+
<div class="daybar"><p>12</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,827: Line 2,022:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>mini-prep of cultivated B0034+ HivC+ilvD+37℃ RBS , B0034+ HivC+ ilvD &amp; HivC(TA) E. coli </p></li>
+
<li class="green"><p>Single colony isolation from mRFP LB-K plate, and cultivation in liquid LB-K</p></li>
-
<li class="green e2"><p>Testing the temperature of the point mutation between of pLac+B0034+Zif268 &amp; AlsS by the m.p 50 &amp; 52℃ of PCR----50℃ is better</p></li>
+
</ul>
</ul>
</div>
</div>
Line 1,834: Line 2,028:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>14</p></div>
+
<div class="daybar"><p>13</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,843: Line 2,041:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>PCR of insert fragment [pLac+B0034+Zif268+AlsS] with designed primer for point mutation -----NOT OK</p></li>
+
<li class="green e5"><p>Ligation: insert[PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1C3]EP and insert[TetR]ES+[B0015]XP/vector[PSB1C3]EP.</p></li>
 +
<li class="green e5"><p>Mini-prep of cultivated mRFP E.coli</p></li>
 +
<li class="green e5"><p>Digestion:[mRFP]ES & [Ter(J61048)]XP & [37OCRBS+mGFP]XP</p></li>
 +
<li class="green e5"><p>Ligation: insert 37oCRBS+LuxR[ES]&37oCRBS+mGFP[XP]  /vector pSB1A3[EP]</p></li>
 +
<li class="green e5"><p>Transformation of 37oCRBS +LuxR+37oCRBS+mGFP on LB-A plate & mRFP +Ter(J61048) on LB-C plate& Ter(J61048) on LB-A plate.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,849: Line 2,051:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>15</p></div>
+
<div class="daybar"><p>14</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,858: Line 2,064:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>DNA Sequencing B0034+ HivC+ilvD+37℃ RBS &amp; B0034+ HivC+ ilvD OK</p></li>
+
<li class="green e5"><p>Transformation of  PompC+B0034+LacI+J61048+PSB1C3 and TetR+B0015+PSB1C3</p></li>
 +
<li class="green e5"><p>Cultivation of B0032+ho1 with liquid LB-A. PCR of insert fragment[Ter(B0015)]</p></li>
 +
<li class="green e5"><p>Single colony isolation from Ter(J61048) LB-A plate, and cultivation in liquid LB-A & 37OCRBS+mGFP on LB-K plate, and cultivation in liquid LB-K</p></li>
 +
<li class="green e5"><p>Transformation of 37oCRBS +LuxR+37oCRBS+mGFP on LB-A plate & mRFP +Ter(J61048) on LB-C plate</p></li>
 +
<li class="green e5"><p>Mini-prep of cultivated Ter (J61048)& 37oCRBS+mGFP E.coli</p></li>
</ul>
</ul>
</div>
</div>
Line 1,864: Line 2,074:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>16</p></div>
+
<div class="daybar"><p>15</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,872: Line 2,086:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>Digestion:[B0032]XP</p></li>
 +
<li class="green e4"><p>Transformation of  PompC+B0034+LacI+J61048 +PSB1C3and TetR+B0015 +PSB1C3 again</p></li>
 +
<li class="green e4"><p>Ligation: insert[PompC]ES+[B0032]XP/vector[PSB1K3]EP, then transformation of this part</p></li>
 +
<li class="green e4"><p>Mini-prep of B0032+ho1. PCR of insert fragment[37oCRBS + LuxR + 37oCRBS+mGFP]&[mRFP+Ter(J61048)].</p></li>
</ul>
</ul>
</div>
</div>
Line 1,879: Line 2,097:
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week">
+
<div class="week bigwk">
<div class="day">
<div class="day">
-
<div class="daybar"><p>17</p></div>
+
<div class="daybar"><p>9</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,889: Line 2,111:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>Mini-prep from cultivated PompC+B0034+LacI+J61048 Eco.li&PSB1C3 E.coli</p></li>
 +
<li class="green e4"><p>digestion : [PompC+B0034+LacI+J61048]ES&[PSB1C3]EP</p></li>
 +
<li class="green e4"><p>Electrophoresis of [PompC+B0034+LacI+J61048] ES& [LacI+J61048]ES&[PSB1C3]EP.</p></li>
 +
<li class="green e4"><p>Single colony isolation from 37oCRBS + mGFP+Ter(B0015)</p></li>
</ul>
</ul>
</div>
</div>
Line 1,894: Line 2,120:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>18</p></div>
+
<div class="daybar"><p>10</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,902: Line 2,131:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e3"><p>Mini-prep of cultivated 37oCRBS+mGFP+Ter(B0015)  E.coli</p></li>
 +
<li class="green e3"><p>Digestion:[ 37oCRBS+mGFP+Ter(B0015)]XP</p></li>
 +
<li class="green e3"><p>Single colony isolation from 37oCRBS+mGFP+Ter(B0015) LB-A plate, and cultivation in liquid LB-A.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,907: Line 2,139:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>19</p></div>
+
<div class="daybar"><p>11</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
Line 1,917: Line 2,149:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [pLac+B0034+Zif268+AlsS] with designed primer for point mutation OK</p></li>
+
<li class="green e2"><p>PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)]</p></li>
-
<li class="green e2"><p>digestion: pLac+B0034+Zif268+AlsS [DPn1]</p></li>
+
<li class="green e2"><p>Transformation of mRFP on LB-K plate.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,924: Line 2,156:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>20</p></div>
+
<div class="daybar"><p>12</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
Line 1,933: Line 2,165:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of pLac+B0034+Zif268+AlsS (point mutation), and cultivation on LB-A plate</p></li>
+
<li class="green"><p>Single colony isolation from mRFP LB-K plate, and cultivation in liquid LB-K</p></li>
</ul>
</ul>
</div>
</div>
Line 1,939: Line 2,171:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>21</p></div>
+
<div class="daybar"><p>13</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,948: Line 2,184:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>Single colony isolation from pLac+B0034+Zif268+AlsS (pm) LB-A plate, and cultivation in liquid LB-A</p></li>
+
<li class="green e5"><p>Ligation: insert[PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1C3]EP and insert[TetR]ES+[B0015]XP/vector[PSB1C3]EP.</p></li>
 +
<li class="green e5"><p>Mini-prep of cultivated mRFP E.coli</p></li>
 +
<li class="green e5"><p>Digestion:[mRFP]ES & [Ter(J61048)]XP & [37OCRBS+mGFP]XP</p></li>
 +
<li class="green e5"><p>Ligation: insert 37oCRBS+LuxR[ES]&37oCRBS+mGFP[XP]  /vector pSB1A3[EP]</p></li>
 +
<li class="green e5"><p>Transformation of 37oCRBS +LuxR+37oCRBS+mGFP on LB-A plate & mRFP +Ter(J61048) on LB-C plate& Ter(J61048) on LB-A plate.</p></li>
</ul>
</ul>
</div>
</div>
Line 1,954: Line 2,194:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>22</p></div>
+
<div class="daybar"><p>14</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 1,964: Line 2,207:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>mini-prep of cultivated pLac+B0034+Zif268+AlsS (pm) E. coli </p></li>
+
<li class="green e5"><p>Transformation of PompC+B0034+LacI+J61048+PSB1C3 and TetR+B0015+PSB1C3</p></li>
-
<li class="green e2"><p>digestion : pLac+B0034+Zif268+AlsS (pm) [EP] </p></li>
+
<li class="green e5"><p>Cultivation of B0032+ho1 with liquid LB-A. PCR of insert fragment[Ter(B0015)]</p></li>
 +
<li class="green e5"><p>Single colony isolation from Ter(J61048) LB-A plate, and cultivation in liquid LB-A & 37OCRBS+mGFP on LB-K plate, and cultivation in liquid LB-K</p></li>
 +
<li class="green e5"><p>Transformation of 37oCRBS +LuxR+37oCRBS+mGFP on LB-A plate & mRFP +Ter(J61048) on LB-C plate</p></li>
 +
<li class="green e5"><p>Mini-prep of cultivated Ter (J61048)& 37oCRBS+mGFP E.coli</p></li>
</ul>
</ul>
</div>
</div>
Line 1,971: Line 2,217:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>23</p></div>
+
<div class="daybar"><p>15</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 1,980: Line 2,229:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>DNA sequencing OK</p></li>
+
<li class="green e4"><p>Digestion:[B0032]XP</p></li>
 +
<li class="green e4"><p>Transformation of  PompC+B0034+LacI+J61048 +PSB1C3and TetR+B0015 +PSB1C3 again</p></li>
 +
<li class="green e4"><p>Ligation: insert[PompC]ES+[B0032]XP/vector[PSB1K3]EP, then transformation of this part</p></li>
 +
<li class="green e4"><p>Mini-prep of B0032+ho1. PCR of insert fragment[37oCRBS + LuxR + 37oCRBS+mGFP]&[mRFP+Ter(J61048)].</p></li>
</ul>
</ul>
</div>
</div>
Line 1,988: Line 2,240:
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week">
+
<div class="week bigwk">
<div class="day">
<div class="day">
-
<div class="daybar"><p>24</p></div>
+
<div class="daybar"><p>16</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 1,998: Line 2,251:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green"><p>PCR + single colony isolation of PompC+B003+LacI+J61048 Cr E.coli&TetR+B0015 Cr E.coli&PompC+B0032 Kr E.coli, then Electrophoresis of  these parts</p></li>
</ul>
</ul>
</div>
</div>
Line 2,003: Line 2,257:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>25</p></div>
+
<div class="daybar"><p>17</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
<li class="caldot green"></li>
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 +
<div class="open">
 +
<ul>
 +
<li class="green"><p>Mini-prep of cultivated PompC+B0034 E.coli and TetR+B0015 Cr E.coli </p></li>
 +
</ul>
 +
</div>
 +
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>18</p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>19</p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>20</p></div>
 +
<div class="dots">
 +
<ul>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,013: Line 2,307:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [B0034+ PBSII+ilvC] with designed primer for point mutation OK</p></li>
+
<li class="green"><p>Ligation: Pcons+RBS+pcyA&B0032+ho1&pSB1A3.</p></li>
-
<li class="green e2"><p>digestion : B0034+ PBSII+ilvC [DPn1]</p></li>
+
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>21</p></div>
 +
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
<li class="green e6"><p>Ligation: insert[PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1A3]EP</p></li>
 +
<li class="green e6"><p>Transformation of Pcons+RBS+pcyA+B0032+ho1 on LB-A plate. PCR of insert fragment[37oCRBS + LuxR + 37oCRBS+mGFP]</p></li>
 +
<li class="green e6"><p>Digestion:[pSB1C3]EP&[mRFP]ES&[Ter(J61048)]XP</p></li>
 +
<li class="green e6"><p>Ligation: insert mRFP[ES]&Ter(J61048)[XP]  /vector pSB1C3[EP]</p></li>
 +
<li class="green e6"><p>Transformation of mRFP + Ter (J61048) on LB-C plate</p></li>
 +
<li class="green e6"><p>Single colony isolation from 37oCRBS + LuxR +37oCRBS+mGFP LB-A plate, and cultivation in liquid LB-A.</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
 +
<div class="day brn">
 +
<div class="daybar"><p>22</p></div>
 +
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 +
<div class="open">
 +
<ul>
 +
<li class="green e3"><p>Transformation of PompC+B0034+LacI+J61048+PSB1A3 but there is no colony appearing. PCR of insert fragment[mRFP+Ter(J61048)]</p></li>
 +
<li class="green e3"><p>Mini-prep of cultivated  37o CRBS + LuxR + 37oCRBS+mGFP E.coli</p></li>
 +
<li class="green e3"><p>Digestion:[ 37o CRBS + LuxR + 37oCRBS+mGFP]ES</p></li>
 +
</ul>
 +
</div>
 +
 +
</div>
 +
</div>
 +
<!---------------------------------------- week end ---------------------------------------->
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week bigwk">
<div class="day">
<div class="day">
-
<div class="daybar"><p>26</p></div>
+
<div class="daybar"><p>23</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,029: Line 2,375:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of B0034+ PBSII+ilvC (pm), and cultivation on LB-C plate------Fail</p></li>
+
<li class="green e6"><p>Digestion:[PompC+B0032]ES and [TetR+B0015]XP,[LacI+J61048]XP</p></li>
 +
<li class="green e6"><p>Ligation: insert [PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1C3]EP</p></li>
 +
<li class="green e6"><p>Transformation  of PompC+B0034+LacI+J61048 +PSB1C3</p></li>
 +
<li class="green e6"><p>Electrophoresis of [PompC+B0032]ES & [TetR+B0015]XP</p></li>
 +
<li class="green e6"><p>Digestion again:[PompC+B0032]ES&[LacI+J61048]XP&[LacI+B0015]XP</p></li>
 +
<li class="green e6"><p>Transformation of mRFP + Ter (J61048) on LB-C plate</p></li>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>24</p></div>
 +
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
<li class="green e7"><p>PCR and single colony isolation of  PompC+B0034+LacI+J61048+PSB1C3 E.coli</p></li>
 +
<li class="green e7"><p>Electrophoresis of [B0032]dig ES&[TetR+B0015]dig XP&[LacI+J61048]dig XP&PCR product [PompC+B0034+LacI+J61048] ,[TetR+B0015] OK</p></li>
 +
<li class="green e7"><p>Decide to determine the sequence of [TetR+B0015] OK.</p></li>
 +
<li class="green e7"><p>Single colony isolation from 6/21 plate.</p></li>
 +
<li class="green e7"><p>Cultivation of Pcons+ RBS+pcyA+B0032+ho1 with liquid LB-A.</p></li>
 +
<li class="green e7"><p>Electrophoresis of Pcons+RBS+PcyA+B0032+ho1.</p></li>
 +
<li class="green e7"><p>PCR of insert fragment[mRFP+Ter(J61048)].</p></li>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>25</p></div>
 +
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
<li class="green e7"><p>Ligation:insert[PompC+B0034]ES+[LacI+J61048]XP&[PompC]ES+[B0032]XP/vector[PSB1C3]EP and [PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1A3]EP</p></li>
 +
<li class="green e7"><p>Transformation of the ligation product made today</p></li>
 +
<li class="green e7"><p>PCR of Pcons+RBS+pcyA+B0032+ho1.</p></li>
 +
<li class="green e7"><p>Cultivation of Pcons+RBS+pcyA+B0032+ho1 with liquid LB-A</p></li>
 +
<li class="green e7"><p>Electrophoresis of Pcons+RBS+pcyA+B0032+ho1.</p></li>
 +
<li class="green e7"><p>Mini-prep of Pcons+RBS+pcyA+B0032+ho1.</p></li>
 +
<li class="green e7"><p>Ligation: insert mRFP[ES]&Ter(J61048)[XP]/vector pSB1C3[EP]</p></li>
</ul>
</ul>
</div>
</div>
Line 2,035: Line 2,440:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>27</p></div>
+
<div class="daybar"><p>26</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,044: Line 2,453:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>PCR of insert fragment [B0034+ PBSII+ilvC] with designed primer for point mutation OK</p></li>
+
<li class="green e5"><p>PCR and single colony isolation of PompC+B0034+LacI+J61048+PSB1A3& PompC+B0034+LacI+J61048+PSB1C3&PompC+B0032+PSB1C3 E.coli</p></li>
 +
<li class="green e5"><p>Electrophoresis of the PCR product made today</p></li>
 +
<li class="green e5"><p>Electrophoresis of Pcons+RBS+pcyA+B0032+ho1──NOT OK</p></li>
 +
<li class="green e5"><p>Ligation: Pcons+RBS+pcyA&B0032+ho1</p></li>
 +
<li class="green e5"><p>Transformation of Pcons+RBS+pcyA+B0032+ho1. Transformation of mRFP + Ter (J61048) on LB-C plate</p></li>
</ul>
</ul>
</div>
</div>
Line 2,050: Line 2,463:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>28</p></div>
+
<div class="daybar"><p>27</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,059: Line 2,476:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>digestion : B0034+ PBSII+ilvC [DPn1]</p></li>
+
<li class="green e5"><p>Cultivation of PompC+B0034+LacI+J61048+PSB1A3 E.coli in liquid LB-A tubes</p></li>
 +
<li class="green e5"><p>PCR of Pcons+RBS+pcyA+B0032+ho1</p></li>
 +
<li class="green e5"><p>Cultivation of Pcons+RBS+pcyA+B0032+ho1 with liquid LB-A</p></li>
 +
<li class="green e5"><p>Electrophoresis of Pcons+RBS+pcyA+B0032+ho1──NOT OK</p></li>
 +
<li class="green e5"><p>Transformation of mRFP + Ter (J61048) on LB-C plate</p></li>
</ul>
</ul>
</div>
</div>
Line 2,065: Line 2,486:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>29</p></div>
+
<div class="daybar"><p>28</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,074: Line 2,497:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of B0034+ PBSII+ilvC (pm), and cultivation on LB-C plate</p></li>
+
<li class="green e3"><p>PCR of Pcons+RBS+pcyA+B0032+ho1</p></li>
 +
<li class="green e3"><p>Electrophoresis of Pcons+RBS+pcAa+B0032+ho1──NOT OK</p></li>
 +
<li class="green e3"><p>Ligation: insert mRFP[ES]&Ter(J61048)[XP]  /vector pSB1C3[EP]</p></li>
</ul>
</ul>
</div>
</div>
Line 2,080: Line 2,505:
</div>
</div>
<div class="day brn">
<div class="day brn">
 +
<div class="daybar"><p>29</p></div>
 +
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 +
<div class="open">
 +
<ul>
 +
<li class="green e8"><p>Cultivation of PompC+B0034+LacI+J61048+PSB1A3 E.coli in liquid LB-A tubes again. </p></li>
 +
<li class="green e8"><p>Ligation: Pcons+RBS+pcyA&B0032+ho1&pSB1A3</p></li>
 +
<li class="green e8"><p>Transformation of ligated Pcons+RBS+pcyA+B0032+ho1</p></li>
 +
<li class="green e8"><p>Single colony isolation from 6/21 Pcons+RBS+pcyA+B0032+ho1 LB plate</p></li>
 +
<li class="green e8"><p>PCR of 6/21 Pcons+RBS+pcyA+B0032+ho1</p></li>
 +
<li class="green e8"><p>Electrophoresis of 621 Pcons+RBS+pcyA+B0032+ho1─NOT OK</p></li>
 +
<li class="green e8"><p>Transformation of mRFP+Ter(J61048) on LB-C plate</p></li>
 +
<li class="green e8"><p>Single colony isolation from 37oCRBS +LuxR +37oCRBS+mGFP LB-A plate, and cultivation in liquid LB-A</p></li>
 +
</ul>
 +
</div>
 +
 +
</div>
 +
</div>
 +
<!---------------------------------------- week end ---------------------------------------->
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week bigwk">
 +
<div class="day">
<div class="daybar"><p>30</p></div>
<div class="daybar"><p>30</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,090: Line 2,554:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [B0034+ PBSII+ilvC] (pm) OK</p></li>
+
<li class="green e8"><p>Mini-prep of cultivated PompC+B0034+LacI+J61048+PSB1A3 E.coli and then digest with EcoR1 and SpeI:[ PompC+B0034+LacI+J61048]dig ES</p></li>
-
<li class="green e2"><p>Single colony isolation from B0034+ PBSII+ilvC (pm) LB-C plate, and cultivation in liquid LB-C</p></li>
+
<li class="green e8"><p>Electrophoresis of [PompC+B0034+LacI+J61048+PSB1A3] mini and [ PompC+B0034+LacI+J61048]dig ES OK</p></li>
 +
<li class="green e8"><p>Decide to determine 
 +
sequence of[ PompC+B0034+LacI+J61048]</p></li>
 +
<li class="green e8"><p>Cultivation of 6/29 Pcons+RBS+PcyA+B0032+ho1 LB plate with liquid LB-A</p></li>
 +
<li class="green e8"><p>PCR of 6/29 Pcons+RBS+PcyA+B0032+ho1</p></li>
 +
<li class="green e8"><p>Cultivation of 6/29 re-ligated Pcons+RBS+PcyA+B0032+ho1</p></li>
 +
<li class="green e8"><p>PCR of 6/29 re-ligated Pcons+RBS+pcyA+B0032+ho1</p></li>
 +
<li class="green e8"><p>Electrophoresis of all above─NOT OK</p></li>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p></p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p></p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p></p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p></p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p></p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day brn">
 +
<div class="daybar"><p></p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
</ul>
</ul>
</div>
</div>
Line 2,120: Line 2,669:
<div class="daysmonth">
<div class="daysmonth">
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week">
+
<div class="week bigwk">
<div class="day">
<div class="day">
-
<div class="daybar"><p>1</p></div>
+
<div class="daybar"><p></p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 2,132: Line 2,679:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>mini-prep of cultivated B0034+ PBSII+ilvC (pm) E. coli </p></li>
 
-
<li class="green e2"><p>digestion : B0034+ PBSII+ilvC (pm) [EP] </p></li>
 
</ul>
</ul>
</div>
</div>
Line 2,139: Line 2,684:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>2</p></div>
+
<div class="daybar"><p>1</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,148: Line 2,697:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>DNA sequencing OK</p></li>
+
<li class="green e5"><p>Ligation: insert [B0030]ES+[ TetR+B0015]XP/vector [PSB1C3]EP </p></li>
 +
<li class="green e5"><p>transformation of this part and cultivation of this part on LB-C plate</p></li>
 +
<li class="green e5"><p>Digestion : mRFP [EP]</p></li>
 +
<li class="green e5"><p>Ligation : insert mRFP [EP]/vector pSB1C3 [EP]</p></li>
 +
<li class="green e5"><p>Transformation of mRFP ,and cultivation on LB-C plate</p></li>
</ul>
</ul>
</div>
</div>
Line 2,154: Line 2,707:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>3</p></div>
+
<div class="daybar"><p>2</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,162: Line 2,717:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e2"><p>PCR +single colony isolation B0030+TetR+B0015  Cr E.coli</p></li>
 +
<li class="green e2"><p>Single colony isolation from mRFP LB-C plate, and cultivation of in liquid LB-C</p></li>
</ul>
</ul>
</div>
</div>
Line 2,167: Line 2,724:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>4</p></div>
+
<div class="daybar"><p>3</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,175: Line 2,739:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e7"><p>Electrophoresis of B0030+TetR+B0015 not OK</p></li>
 +
<li class="green e7"><p>electrophoresis  of B0030+TetR+B0015  again OK</p></li>
 +
<li class="green e7"><p>Ligation: Pcons+B0030+pcyA+B0032+ho1</p></li>
 +
<li class="green e7"><p>Transformation of Pcons+B0030+pcyA+B0032+ho1</p></li>
 +
<li class="green e7"><p>Mini-prep of cultivated 37℃RBS+luxR+37℃RBS+mGFP and mRFP E.coli</p></li>
 +
<li class="green e7"><p>Digestion : mRFP [ES] & pSB1A3 [EP] & pSB1C3 [EP] & pSB1K3 [EP]</p></li>
 +
<li class="green e7"><p>Electrophoresis of insert fragment [mRFP & pSB1A3 & pSB1C3 & pSB1K3]---- OK</p></li>
</ul>
</ul>
</div>
</div>
Line 2,180: Line 2,751:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>5</p></div>
+
<div class="daybar"><p>4</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,188: Line 2,764:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e5"><p>Cultivation of B0030+TetR+B0015+PSB1C3 in liquid LB-C tubes. PCR  of  Pcons+B0030+pcyA+B0032+ho1</p></li>
 +
<li class="green e5"><p>Electrophoresis of Pcons+B0030+pcyA+B0032+ho1</p></li>
 +
<li class="green e5"><p>Cultivation of Pcons+B0030+pcyA+B0032+ho1 with liquid LB-A</p></li>
 +
<li class="green e5"><p>Ligation : insert mRFP [ES] & J61048 [XP]/ vector pSB1K3 [EP]</p></li>
 +
<li class="green e5"><p>Transformation of mRFP+J61048 ,and cultivation on LB-K plate</p></li>
</ul>
</ul>
</div>
</div>
Line 2,193: Line 2,774:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>6</p></div>
+
<div class="daybar"><p>5</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,201: Line 2,788:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e6"><p>Mini-prep of cultivated B0030+TetR+B0015+PSB1C3 Cr E.coli and then digest with XbaI and Pst1</p></li>
 +
<li class="green e6"><p>Electrophoresis  of[ B0030+TetR+B0015]mini&dig XP
 +
</p></li>
 +
<li class="green e6"><p>Decide to determine the sequence of B0030+TetR+B0015</p></li>
 +
<li class="green e6"><p>Digestion: [Pcons+B0030+pcyA+B0032+ho1]ES</p></li>
 +
<li class="green e6"><p>Electrophoresis of Pcons+B0030+pcyA+B0032+ho1──NOT OK</p></li>
 +
<li class="green e6"><p>PCR of insert fragment [mRFP+J61048]</p></li>
</ul>
</ul>
</div>
</div>
Line 2,206: Line 2,800:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>7</p></div>
+
<div class="daybar"><p>6</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,215: Line 2,813:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>digestion : pLac+B0034+zif268+AlsS (pm) [ES] &amp; B0034+ PBSII+ilvC (pm) [XP]</p></li>
+
<li class="green e5"><p>Ligation :insert [PompC]ES+[B0032]XP/vector [PSB1C3] EP and insert [LacI+J61048]ES&[Plac]XP/vector [PSB1C3]EP</p></li>
 +
<li class="green e5"><p>Transformation of PompC+B0030+PSB1C3 and LacI+J61048+Plac+PSB1C3</p></li>
 +
<li class="green e5"><p>Ligation : insert 37℃RBS+luxR+37℃RBS+mGFP [ES] & B0015 [XP]/ vector pSB1C3 [EP]</p></li>
 +
<li class="green e5"><p>Transformation of 37℃RBS+luxR+37℃RBS+mGFP+B0015 ,and cultivation on LB-C plate</p></li>
 +
<li class="green e5"><p>Electrophoresis of insert fragment [mRFP+J61048]----undetermined</p></li>
</ul>
</ul>
</div>
</div>
Line 2,224: Line 2,826:
<div class="week mweek">
<div class="week mweek">
<div class="day">
<div class="day">
-
<div class="daybar"><p>8</p></div>
+
<div class="daybar"><p>7</p></div>
<div class="dots">
<div class="dots">
 +
<ul>
 +
</ul>
</div>
</div>
<div class="open">
<div class="open">
 +
<ul>
 +
</ul>
</div>
</div>
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>9</p></div>
+
<div class="daybar"><p>8</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,245: Line 2,849:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e4"><p>Testing the temperature of the point mutation between of HivC &amp; ilvD by the m.p 50℃ of PCR</p></li>
+
<li class="green e2"><p>PCR + single colony isolation of LacI+J61048+Plac +PSB1C3 E.coli</p></li>
-
<li class="green e4"><p>digestion : pSB1K3 [EP]</p></li>
+
<li class="green e2"><p>Electrophoresis of LacI+J61048+Plac OK</p></li>
-
<li class="green e4"><p>ligation : insert pLac+B0034+zif268+AlsS (pm) [ES] &amp; B0034+ PBSII+ilvC (pm) [XP]/vector pSB1K3 [EP]</p></li>
+
-
<li class="green e4"><p>transformation of pLac+B0034+zif268+AlsS+ B0034+ PBSII+ilvC (pm) and cultivation on LB-K plate</p></li>
+
</ul>
</ul>
</div>
</div>
Line 2,254: Line 2,856:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>10</p></div>
+
<div class="daybar"><p>9</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,266: Line 2,867:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e4"><p>Testing the temperature of the point mutation between of HivC &amp; ilvD by the m.p 48℃ of PCR</p></li>
+
<li class="green e3"><p>Single colony isolation from mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K</p></li>
-
<li class="green e4"><p>Digestion : B0034+ HivC+ilvD+37℃ RBS [DPn1] </p></li>
+
<li class="green e3"><p>Mini-prep of cultivated mRFP+J61048 E.coli</p></li>
-
<li class="green e4"><p>PCR of insert fragment [pLac+B0034+zif268+AlsS+ B0034+ PBSII+ilvC] (pm) OK</p></li>
+
<li class="green e3"><p>Digestion : mRFP+J61048 [XP]</p></li>
-
<li class="green e4"><p>SCI from pLac+B0034+zif268+AlsS+ B0034+ PBSII+ilvC (pm) LB-K plate, and cultivation in liquid LB-K</p></li>
+
</ul>
</ul>
</div>
</div>
Line 2,275: Line 2,875:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>11</p></div>
+
<div class="daybar"><p>10</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,286: Line 2,885:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>mini-prep of cultivated pLac+B0034+zif268+AlsS+ B0034+ PBSII+ilvC (pm)  E. coli </p></li>
+
<li class="green e2"><p>Cultivation
-
<li class="green e3"><p>DNA sequencing OK</p></li>
+
of LacI+J61048+Plac in liquid LB-C tubes.</p></li>
-
<li class="green e3"><p>culture condition test: activation DH5αovernight</p></li>
+
<li class="green e2"><p>Electrophoresis of insert fragment [mRFP+J61048]----NOT OK</p></li>
</ul>
</ul>
</div>
</div>
Line 2,294: Line 2,893:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>12</p></div>
+
<div class="daybar"><p>11</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,304: Line 2,902:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>transfer to new medium(1/100), OD0.2 start counting culture time</p></li>
+
<li class="green"><p>Mini-prep of cultivated LacI+J61048+Plac E.coli</p></li>
-
<li class="green e2"><p>transfer to 30゜C and 27゜C</p></li>
+
</ul>
</ul>
</div>
</div>
Line 2,311: Line 2,908:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>13</p></div>
+
<div class="daybar"><p>12</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,320: Line 2,923:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transfer to 30゜C and 27゜C</p></li>
+
<li class="green e7"><p>Ligation again:Insert [PompC]ES&[B0032]XP/vector [PSB1K3]EP</p></li>
 +
<li class="green e7"><p>Digestion: [LacI+J61048+Plac]XP</p></li>
 +
<li class="green e7"><p>Electrophoresis of [LacI+J61048+Plac]mini&dig XP OK</p></li>
 +
<li class="green e7"><p>Decide to determine the sequence of LacI+J61048+Plac</p></li>
 +
<li class="green e7"><p>Transformation of PompC(Ar)+B0032(Ar)+PSB1K3 again but there is still no colony</p></li>
 +
<li class="green e7"><p>Transformation of 37℃RBS+luxR+37℃RBS+mGFP+B0015, and cultivation on LB-C plate</p></li>
 +
<li class="green e7"><p>Transformation of mRFP+J61048 ,and cultivation on LB-K plate.</p></li>
</ul>
</ul>
</div>
</div>
Line 2,326: Line 2,935:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>14</p></div>
+
<div class="daybar"><p>13</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,334: Line 2,944:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green"><p>Ligation: Insert [PompC]ES&[B0032]XP/vector [PSB1K3]EP. PCR of insert fragment [37℃RBS+luxR+37℃RBS+mGFP+B0015 & mRFP+J61048].</p></li>
</ul>
</ul>
</div>
</div>
Line 2,342: Line 2,953:
<div class="week mweek">
<div class="week mweek">
<div class="day">
<div class="day">
-
<div class="daybar"><p>15</p></div>
+
<div class="daybar"><p>14</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,351: Line 2,964:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of B0034+ HivC+ ilvD+37℃ RBS (pm),and cultivation on LB-A plate</p></li>
+
<li class="green e3"><p>Transformation of the ligation product made yesterday on LB-K plate  but there is no colony appearing</p></li>
 +
<li class="green e3"><p>Electrophoresis of insert fragment [37℃RBS+luxR+37℃RBS+mGFP+B0015]----OK</p></li>
 +
<li class="green e3"><p>Electrophoresis of insert fragment [mRFP+J61048]----NOT OK
 +
RBS+mGFP+B0015 LB-C plate, and cultivation of in liquid LB-C. PCR of insert fragment [mRFP+J61048]----NOT OK</p></li>
</ul>
</ul>
</div>
</div>
Line 2,357: Line 2,973:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>16</p></div>
+
<div class="daybar"><p>15</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,366: Line 2,989:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>PCR of insert fragment [B0034+ HivC+ilvD+37℃ RBS](pm)-----NOT OK</p></li>
+
<li class="green e8"><p>Ligation and trans formation of insert [ B0030]ES+ [TetR+B0015]XP/vector [PSB1C3]EP  and cultivation on LB-C plate</p></li>
 +
<li class="green e8"><p>Transformation of PSB1K3 for testing</p></li>
 +
<li class="green e8"><p>Transformation of  Pcons</p></li>
 +
<li class="green e8"><p>Digestion: [B0030+pcyA]ES</p></li>
 +
<li class="green e8"><p>Electrophoresis of Pcons──OK</p></li>
 +
<li class="green e8"><p>Electrophoresis of insert fragment [mRFP+J61048]----NOT OK</p></li>
 +
<li class="green e8"><p>Mini-prep of cultivated 37℃RBS+luxR+37℃RBS+mGFP+B0015 E.coli----NOT OK</p></li>
 +
<li class="green e8"><p>PCR of insert fragment [mRFP+J61048]----OK</p></li>
</ul>
</ul>
</div>
</div>
Line 2,372: Line 3,002:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>17</p></div>
+
<div class="daybar"><p>16</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,382: Line 3,018:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Point mutation by PCR of insert fragment [B0034+ HivC+ilvD+37℃ RBS]</p></li>
+
<li class="green e8"><p>Digestion:[PompC]ES,[PSB1K3]&[PSB1C3]EPand Electrophoresis for checking these parts OK</p></li>
-
<li class="green e2"><p>Digestion: B0034+ HivC+ilvD+37℃ RBS [DPn1] </p></li>
+
<li class="green e8"><p>PCR + singles colony isolation of B0030+TetR+B0015 +PSB1C3 E.coli and electrophoresis of this insert fragment not OK Because the PCR electrophoresis result is strange，we conduct this experience again OK</p></li>
 +
<li class="green e8"><p>Ligation: Pcons&Pcons+B0030+pcyA+B0032+ho1&pSB1K3.</p></li>
 +
<li class="green e8"><p>Transformation of Pcons+B0032+pcyA+B0030+ho1.</p></li>
 +
<li class="green e8"><p>Cultivation of Pcons with liquid LB-C</p></li>
 +
<li class="green e8"><p>Electrophoresis of Pcons──NOT OK</p></li>
 +
<li class="green e8"><p>Single colony isolation from 37℃RBS+luxR+37℃RBS+mGFP+B0015 LB-C plate, and cultivation of in liquid LB-C</p></li>
 +
<li class="green e8"><p>Single colony isolation from mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K</p></li>
</ul>
</ul>
</div>
</div>
Line 2,389: Line 3,031:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>18</p></div>
+
<div class="daybar"><p>17</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,397: Line 3,047:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e8"><p>Transformation of PompC+B0032+PSB1C3 and
 +
Cultivation of B0030+TetR+B0015 Cr E.coli  in  liquid LB-C tube. </p></li>
 +
<li class="green e8"><p>Transformation of Pcons</p></li>
 +
<li class="green e8"><p>transformation of tetR plasmid and cultivation on LB –A plate  3. Digestion: ter. [J61048], ter. [B0015] dig EP</p></li>
 +
<li class="green e8"><p>Digestion : pSB1C3 [ES] & pSB1C3 [XP]</p></li>
 +
<li class="green e8"><p>Ligation : insert sRNA+target 1 [ES]/vector pSB1C3 [ES]</p></li>
 +
<li class="green e8"><p>Transformation of 37℃RBS+luxR+37℃RBS+mGFP+B0015 & sRNA 1 & sRNA 2 & target 1 & target 2 ,and cultivation on LB-C plate</p></li>
 +
<li class="green e8"><p>Transformation of mRFP+J61048 & sRNA+target 1 & sRNA+target 2 ,and cultivation on LB-K plate</p></li>
 +
<li class="green e8"><p>Electrophoresis of insert fragment [37℃RBS+luxR+37℃RBS+mGFP+B0015 & mRFP+J61048 & pSB1A3 & pSB1C3 & pSB1K3]-- mRFP+J61048 NOT OK Digestion : mRFP+J61048 [XP]</p></li>
</ul>
</ul>
</div>
</div>
Line 2,402: Line 3,061:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>19</p></div>
+
<div class="daybar"><p>18</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,411: Line 3,077:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of B0034+ HivC+ ilvD+37℃ RBS (pm),and cultivation on LB-A,K,C plate (A sucessful)</p></li>
+
<li class="green e8"><p>Mini-prep of cultivated B0030+TetR+B0015 Cr E.coli and then digest these mini with XbaI and PstI</p></li>
 +
<li class="green e8"><p>PCR + single colony isolation of PompC+B0032 Cr E.coli and electrophoresis of this  insert fragment OK Cultivation of [PompC+B0032] Cr  E.coli in liquid LB-C Tubes.</p></li>
 +
<li class="green e8"><p>Cultivation of Pcons with liquid LB-C</p></li>
 +
<li class="green e8"><p>Digestion : pSB1C3 [ES] & pSB1C3 [XP]</p></li>
 +
<li class="green e8"><p>Mini-prep of Pcons. Electrophoresis of Pcons──NOT OK</p></li>
 +
<li class="green e8"><p>Four colonies isolation from Pcons LB plate and use streak plate method to isolate the pure cultures. Digestion : mRFP+J61048 [XP]</p></li>
 +
<li class="green e8"><p>Single colony isolation from sRNA+target 1 & sRNA+target 2 LB-K plate, and cultivation of in liquid LB-K</p></li>
 +
<li class="green e8"><p>Single colony isolation from Plux LB-A plate, and cultivation of in liquid LB-A----NOT OK</p></li>
</ul>
</ul>
</div>
</div>
Line 2,417: Line 3,090:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>20</p></div>
+
<div class="daybar"><p>19</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,427: Line 3,106:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [B0034+ HivC+ilvD+37℃ RBS](pm) ---- OK</p></li>
+
<li class="green e8"><p>Mini-prep of cultivated PompC+B0032 Cr E.coli, And then digest with EcoR1 and SpeI</p></li>
-
<li class="green e2"><p>Single colony isolation from B0034+ HivC+ilvD+37℃ RBS (pm) LB-A plate, and cultivation in liquid LB-A</p></li>
+
<li class="green e8"><p>Electrophoresis of PSB1K3 made at 7/16 and B0030+TetR+B0015, PompC+B0032 OK.</p></li>
 +
<li class="green e8"><p>Cultivation of Pcons with liquid LB-C</p></li>
 +
<li class="green e8"><p>Single colony isolation from37℃RBS+luxR+37℃RBS+mGFP+B0015 LB-C plate, and cultivation of in liquid LB-C</p></li>
 +
<li class="green e8"><p>Single colony isolation from mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K(?)</p></li>
 +
<li class="green e8"><p>PCR of insert fragment [sRNA 1 & sRNA 2 & target 1 & target 2]</p></li>
 +
<li class="green e8"><p>Electrophoresis of insert fragment [sRNA 1 & sRNA 2 & target 1 & target 2]---- target 1?</p></li>
 +
<li class="green e8"><p>Mini-prep of cultivated sRNA+target 1 & sRNA+target 2 E.coli.</p></li>
</ul>
</ul>
</div>
</div>
Line 2,434: Line 3,119:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>21</p></div>
+
<div class="daybar"><p>20</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,445: Line 3,135:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>mini-prep of cultivated B0034+ HivC+ilvD+37℃ RBS (pm) E. coli </p></li>
+
<li class="green e8"><p>Mini-prep of Pcons.
-
<li class="green e3"><p>Digestion: pLac+B0034+zif268+AlsS+ B0034+ PBSII+ilvC (pm)(G1) [ES] &amp; B0034+ HivC+ilvD+37℃ RBS (pm)(G2) [XP] </p></li>
+
-Digestion: [Pcons]ES.</p></li>
-
<li class="green e3"><p>ligation : insert G1[ES] &amp; G2[XP]
+
<li class="green e8"><p>Electrophoresis of Pcons──NOT OK.</p></li>
-
          vector pSB1C3[EP]
+
<li class="green e8"><p>Single colony isolation from sRNA 1 & sRNA 2 & target 2 LB-C plate, and cultivation of in liquid LB-C</p></li>
-
</p></li>
+
<li class="green e8"><p>Single colony isolation from mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K</p></li>
 +
<li class="green e8"><p>Transformation of Plux ,and cultivation on LB-A plate</p></li>
 +
<li class="green e8"><p>Single colony isolation from Plux & PLac & Pcons LB-A plate, and cultivation of in liquid LB-A</p></li>
 +
<li class="green e8"><p>Transformation of RBS+lacI+B0010+B0012+pLac ,and cultivation on LB-K plate</p></li>
 +
<li class="green e8"><p>Mini-prep of cultivated 37℃RBS+luxR+37℃RBS+mGFP+B0015 & mRFP+J61048 & sRNA 1 & sRNA 2 & target 2 E.coli</p></li>
</ul>
</ul>
</div>
</div>
Line 2,457: Line 3,151:
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week">
+
<div class="week bigwk">
<div class="day">
<div class="day">
-
<div class="daybar"><p>22</p></div>
+
<div class="daybar"><p>21</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,468: Line 3,166:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of G1+G2,and cultivation on LB- C plate failed</p></li>
+
<li class="green e5"><p>Single colony isolation from 7/15 Pcons LB plate(Red one).</p></li>
 +
<li class="green e5"><p>Cultivation of Pcons with LB-C. PCR of insert fragment [target 1]</p></li>
 +
<li class="green e5"><p>Transformation of RBS+lacI+B0010+B0012+pLac ,and cultivation on LB-K plate</p></li>
 +
<li class="green e5"><p>Electrophoresis of insert fragment [37℃RBS+luxR+37℃RBS+mGFP+B0015 & mRFP+J61048 & sRNA 1 & sRNA 2 & target 1 & target 2]----?</p></li>
 +
<li class="green e5"><p>Mini-prep of cultivated Plux & PLac E.coli
 +
Digestion : Plux [ES] & Plac [ES] & mRFP+J61048 [XP].</p></li>
</ul>
</ul>
</div>
</div>
Line 2,474: Line 3,177:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>23</p></div>
+
<div class="daybar"><p>22</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,483: Line 3,193:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>ligation : insert G1[ES] &amp; G2[XP]/vector pSB1C3[EP]</p></li>
+
<li class="green e8"><p>Ligation :insert [LacI+J61048]ES&[Plac]XP/vector [PSB1C3] EP</p></li>
 +
<li class="green e8"><p>Transformation of the ligation product made at this morning and cultivation on LB-C plate.</p></li>
 +
<li class="green e8"><p>Mini-prep of Pcons+mRFP</p></li>
 +
<li class="green e8"><p>Digestion: [Pcons]ES.</p></li>
 +
<li class="green e8"><p>Electrophoresis of Pcons+mRFP──NOT OK.</p></li>
 +
<li class="green e8"><p>Cultivation of Pcons+mRFP with liquid LB-C. PCR of Pcons+mRFP</p></li>
 +
<li class="green e8"><p>Electrophoresis of Pcons+mRFP──OK.</p></li>
 +
<li class="green e8"><p>Electrophoresis of insert fragment [Plux & Plac & mRFP+J61048?]----?</p></li>
</ul>
</ul>
</div>
</div>
Line 2,489: Line 3,206:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>24</p></div>
+
<div class="daybar"><p>23</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,500: Line 3,222:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>transformation of G1+G2,and cultivation on LB- C plate</p></li>
+
<li class="green e8"><p>PCR +Single colony isolation  of   LacI+J61048+Plac and electrophoresis Of  this  insert fragment  OK</p></li>
-
<li class="green e3"><p>Digestion: G2’(B0034+HIVC+ilvD) [XP]</p></li>
+
<li class="green e8"><p>Mini-prep of Pcons-mRFP.</p></li>
-
<li class="green e3"><p>ligation : insert G1[ES] &amp; G2‘[XP]/vector pSB1C3[EP]</p></li>
+
<li class="green e8"><p>Digestion: [B0030]XP&[B0032]XP&[B0023]XP&[Pcons]ES</p></li>
 +
<li class="green e8"><p>Electrophoresis of B0030,B0032,B0034,Pcons──OK</p></li>
 +
<li class="green e8"><p>Transformation of B0030+pcyA+B0032+ho1+B0030/B0032/B0034</p></li>
 +
<li class="green e8"><p>Electrophoresis of insert fragment [mRFP+J61048]----?</p></li>
 +
<li class="green e8"><p>Transformation of RBS+lacI+B0010+B0012+Plac ,and cultivation on LB-K plate----NOT OK</p></li>
 +
<li class="green e8"><p>Single colony isolation from RBS+lacI+B0010+B0012+pLac & mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K</p></li>
</ul>
</ul>
</div>
</div>
Line 2,508: Line 3,235:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>25</p></div>
+
<div class="daybar"><p>24</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,518: Line 3,251:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [kivD+B0015]</p></li>
+
<li class="green e8"><p>Cultivation of LacI+J61048+Plac E.coli in liquid LB-C tubes</p></li>
-
<li class="green e2"><p>transformation of G1+G2’, and cultivation on LB- C plate failed</p></li>
+
<li class="green e8"><p>Mini-prep of cultivated LacI+J61048+Plac E.coli. </p></li>
 +
<li class="green e8"><p>Cultivation of B0030+pcyA+B0032+ho1+B0032 with liquid LB</p></li>
 +
<li class="green e8"><p>Transformation of B0030+pcyA+B0032+ho1+B0030/B0032/B0034</p></li>
 +
<li class="green e8"><p>Mini-prep of cultivated mRFP+J61048 E.coli</p></li>
 +
<li class="green e8"><p>Digestion : mRFP+J61048 [XP]</p></li>
 +
<li class="green e8"><p>Transformation of target 1 ,and cultivation on LB-C plate</p></li>
 +
<li class="green e8"><p>Single colony isolation from RBS+lacI+B0010+B0012+pLac plate, and cultivation of in liquid LB-without antibiotics----NO E.coli?</p></li>
</ul>
</ul>
</div>
</div>
Line 2,525: Line 3,264:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>26</p></div>
+
<div class="daybar"><p>25</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,535: Line 3,280:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Single colony isolation from kivD+B0015 LB-C plate, and cultivation in liquid LB-C</p></li>
+
<li class="green e8"><p>Digestion of LacI+J61048+Plac Cr [XP]</p></li>
-
<li class="green e2"><p>sample and do GC</p></li>
+
<li class="green e8"><p>Electrophoresis of[ LacI+J61048 +Plac]dig XP  OK</p></li>
 +
<li class="green e8"><p>Decide to determine the sequence of  LacI+J61048 +Plac</p></li>
 +
<li class="green e8"><p>Ligation :insert [B0030]ES& [TetR+B0015]XP/vector [PSB1K3]EP.</p></li>
 +
<li class="green e8"><p>Cultivation of B0030+pcyA+B0032+ho1+B0030/B0032/B0034 with liquid LB.</p></li>
 +
<li class="green e8"><p>PCR of insert fragment [target 1]</p></li>
 +
<li class="green e8"><p>Single colony isolation from RBS+lacI+B0010+B0012+pLac plate , and cultivation of in liquid LB-without antibiotics----NO E.coli?</p></li>
 +
<li class="green e8"><p>Electrophoresis of insert fragment [mRFP+J61048 & target 1]----?</p></li>
</ul>
</ul>
</div>
</div>
Line 2,542: Line 3,293:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>27</p></div>
+
<div class="daybar"><p>26</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,551: Line 3,308:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>mini-prep of cultivated kivD+B0015 E. coli </p></li>
+
<li class="green e7"><p>Transformation of B0030+TetR+B0015+PSB1K3 and cultivation on LB-K plate. -Mini-prep of B0030+pcyAB0032+ho1+B0030/B0032/B0034.</p></li>
 +
<li class="green e7"><p>Digestion: [B0030+pcyA+B0032+ho1+B0030/B0032/B0034]XP</p></li>
 +
<li class="green e7"><p>Electrophoresis of digested B0030+pcyA+B0032+ho1+B0030/B0032/B0034──OK</p></li>
 +
<li class="green e7"><p>Electrophoresis of B0030,B0032,B0034,Pcons──OK</p></li>
 +
<li class="green e7"><p>Mini-prep of cultivated target 1 & mRFP+J61048 E.coli</p></li>
 +
<li class="green e7"><p>Digestion: target 1 [ES] & mRFP [XP]?</p></li>
 +
<li class="green e7"><p>Electrophoresis of insert fragment [RBS+lacI+B0010+B0012+pLac & mRFP & target 1]----?</p></li>
</ul>
</ul>
</div>
</div>
Line 2,557: Line 3,320:
</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p>28</p></div>
+
<div class="daybar"><p>27</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,565: Line 3,331:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e3"><p>PCR and single colony isolation of B0030+TetR+B0015 Kr E.coli</p></li>
 +
<li class="green e3"><p>Cultivation of B0030+TetR+B0015 in liquid LB-tube.</p></li>
 +
<li class="green e3"><p>Ligation: pSB1C3&Pcons&B0030+pcyA+B0032+ho1+B0030</p></li>
</ul>
</ul>
</div>
</div>
Line 2,572: Line 3,341:
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week end ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
-
<div class="week">
+
<div class="week mweek">
<div class="day">
<div class="day">
-
<div class="daybar"><p>29</p></div>
+
<div class="daybar"><p>28</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,584: Line 3,355:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Point mutation by PCR of insert fragment [B0034+ HivC+ilvD(G2’)]</p></li>
+
<li class="green e4"><p>Cultivation of B0030+TetR+B0015 in liquid LB-K tubes again</p></li>
-
<li class="green e2"><p>Digestion: G2’ [DPn1]</p></li>
+
<li class="green e4"><p>Ligation:insert[PompC+B0032]ES+[LacI+J61048+Plac]XP/vector[PSB1K3]EP</p></li>
 +
<li class="green e4"><p>Mini-prep of cultivated B0030+TetR+B0015 Kr E.coli</p></li>
 +
<li class="green e4"><p>Transformation of Pcons+B0030+pcyA+B0032+ho1+B0030</p></li>
</ul>
</ul>
</div>
</div>
Line 2,591: Line 3,364:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>30</p></div>
+
<div class="daybar"><p>29</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,600: Line 3,374:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transformation of G2’, and cultivation on LB- A plate</p></li>
+
<li class="green e2"><p>Transformation of PompC+B0032+LacI+J61048+Plac+PSB1K3. -Transformation of Cph8.</p></li>
 +
<li class="green e2"><p>Digestion: [Pcons]XP.</p></li>
</ul>
</ul>
</div>
</div>
Line 2,606: Line 3,381:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p>31</p></div>
+
<div class="daybar"><p>30</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,616: Line 3,392:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Digestion: kivD+B0015 [EP] (checking bp----failed)</p></li>
+
<li class="green e3"><p>Ligation:insert[PompC+B0032]ES+[LacI+J61048+Plac]XP/vector[PSB1K3]EP&[PSB1A3]EP</p></li>
-
<li class="green e2"><p>Single colony isolation from G2’ (pm) LB-A plate, and cultivation in liquid LB-A</p></li>
+
<li class="green e3"><p>Transformation of PompC+B0032+LacI+J61048+Plac+PSB1K3&PSB1A3.</p></li>
 +
<li class="green e3"><p>Electrophoresis of Pcons ─OK. Digestion: [Pcons]SP.</p></li>
</ul>
</ul>
</div>
</div>
Line 2,623: Line 3,400:
</div>
</div>
<div class="day">
<div class="day">
-
<div class="daybar"><p></p></div>
+
<div class="daybar"><p>31</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,631: Line 3,412:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>PCR + single colony isolation of PompC+B0032+LacI+J61048+Plac+PSB1K3&PSB1A3 E.coli</p></li>
 +
<li class="green e4"><p>Electrophoresis of PompC+B0032+LacI+J61048+Plac OK.</p></li>
 +
<li class="green e4"><p>Ligation: Pcons&B0030+pcyA+B0032+ho1+B0030.</p></li>
 +
<li class="green e4"><p>Transformation of Pcons+B0030+pcyA+B0032+ho1+B0030</p></li>
</ul>
</ul>
</div>
</div>
Line 2,699: Line 3,484:
<!---------------------------------------- week start ---------------------------------------->
<!---------------------------------------- week start ---------------------------------------->
<div class="week">
<div class="week">
 +
<div class="day">
 +
<div class="daybar"><p></p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 +
</div>
<div class="day">
<div class="day">
<div class="daybar"><p></p></div>
<div class="daybar"><p></p></div>
Line 2,742: Line 3,540:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,748: Line 3,547:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>mini-prep of cultivated G2’ E. coli</p></li>
+
<li class="green e2"><p>Cultivation of PompC+B0032+LacI+J61048+Plac Kr&Ar E.coli in liquid LB-K&LB-A tubes.</p></li>
 +
<li class="green e2"><p>Cultivation of Pcons+B0030+pcyA+ B0032+ho1+B0030 with liquid LB-K</p></li>
</ul>
</ul>
</div>
</div>
Line 2,757: Line 3,557:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,762: Line 3,566:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>Mini-prep of cultivated PompC+B0032+LacI+J61048+Plac Kr&Ar E.coli</p></li>
 +
<li class="green e4"><p>Digestion: [PompC+B0032+LacI+J61048]ES. Mini-prep of Pcons+B0030+pcyA+ B0032+ho1+B0030.</p></li>
 +
<li class="green e4"><p>Digestion: [Pcons+B0030+pcyA+ B0032+ho1+B0030]ES</p></li>
 +
<li class="green e4"><p>Electrophoresis of Pcons+B0030+pcyA+ B0032+ho1+B0030──OK</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
+
<div class="day brn">
<div class="daybar"><p>3</p></div>
<div class="daybar"><p>3</p></div>
<div class="dots">
<div class="dots">
Line 2,778: Line 3,586:
</div>
</div>
-
</div>
+
</div>
-
<div class="day brn">
+
</div>
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week">
 +
<div class="day">
<div class="daybar"><p>4</p></div>
<div class="daybar"><p>4</p></div>
<div class="dots">
<div class="dots">
Line 2,791: Line 3,602:
</div>
</div>
-
</div>
+
</div>
-
</div>
+
-
<!---------------------------------------- week start ---------------------------------------->
+
-
<div class="week">
+
<div class="day">
<div class="day">
<div class="daybar"><p>5</p></div>
<div class="daybar"><p>5</p></div>
Line 2,849: Line 3,657:
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,856: Line 3,663:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Digestion: G1 [ES] &amp; G2‘ [XP] &amp; pSB1C3 [EP]</p></li>
+
<li class="green"><p>Ligation: insert[PompC+B0032]ES+[LacI+J61048+Plac]XP/vector[PSB1K3]EP, then transformation of this ligation product</p></li>
-
<li class="green e2"><p>ligation : insert G1[ES] &amp; G2‘[XP]/vector pSB1C3[EP]</p></li>
+
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
+
<div class="day brn">
<div class="daybar"><p>10</p></div>
<div class="daybar"><p>10</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
-
<li class="caldot green"></li>
 
</ul>
</ul>
</div>
</div>
Line 2,872: Line 3,677:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>strain test: activation of different strains overnight</p></li>
 
</ul>
</ul>
</div>
</div>
-
</div>
+
</div>
-
<div class="day brn">
+
</div>
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week bigwk">
 +
<div class="day">
<div class="daybar"><p>11</p></div>
<div class="daybar"><p>11</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,887: Line 3,695:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transfer to new medium, OD0.2 IPTGinduction, culture in 37゜C</p></li>
+
<li class="green e2"><p>PCR + single colony isolation of PompC+B0032+LacI+J61048+Plac Kr E.coli</p></li>
 +
<li class="green e2"><p>Electrophoresis of PompC+B0032+LacI+J61048+Plac not OK</p></li>
</ul>
</ul>
</div>
</div>
-
</div>
+
</div>
-
</div>
+
-
<!---------------------------------------- week start ---------------------------------------->
+
-
<div class="week">
+
<div class="day">
<div class="day">
<div class="daybar"><p>12</p></div>
<div class="daybar"><p>12</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,906: Line 3,713:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Single colony isolation from G1+G2’ LB-C plate, and cultivation in liquid LB-C</p></li>
+
<li class="green e3"><p>Colony PCR of 37。C RBS+LuxR+37。C RBS+mGFP Ar E.coli&Plux+sRNA2 Kr E.coli&mRFP+J61048 Kr E.coli</p></li>
-
<li class="green e2"><p>transfer to 27゜C</p></li>
+
<li class="green e3"><p>Electrophoresis of [target site 1]PCR&[37。C RBS+LuxR+37。C RBS+mGFP]PCR&[LacI+J61048+Plac+sRNA2]dig XP&[target site2+mRFP]dig ES&[Plux+sRNA2]PCR&[mRFP+J61048]PCR</p></li>
 +
<li class="green e3"><p>Cutivation of Plux+sRNA2 Kr E.coli in liquid LB-K tube</p></li>
</ul>
</ul>
</div>
</div>
Line 2,916: Line 3,724:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 2,922: Line 3,733:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>mini-prep of cultivated G1+G2’ E. coli </p></li>
+
<li class="green e4"><p>Mini-prep of cultivated Plux+sRNA2 Kr E.coli(fault)</p></li>
 +
<li class="green e4"><p>Transformation of 37。C RBS+LuxR+37。C RBS+mGFP in PSB1A3</p></li>
 +
<li class="green e4"><p>Ligation:insert[sRNA+target site1]XP/vector[PSB1C3]XP Colony PCR of target site2+mRFP Ar E.coli</p></li>
 +
<li class="green e4"><p>Cultivation of Plux+sRNA2 Kr E.coli in liquid LB-K tube</p></li>
</ul>
</ul>
</div>
</div>
Line 2,931: Line 3,745:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 2,936: Line 3,757:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e7"><p>Mini-prep of cultivated Plux+sRNA2 Kr E.coli</p></li>
 +
<li class="green e7"><p>Digestion:[Plux+sRNA2]XP</p></li>
 +
<li class="green e7"><p>Digestion:[Plux+sRNA2]XP</p></li>
 +
<li class="green e7"><p>Colony PCR of 37。C RBS+LuxR+37。C RBS+mGFP Ar E.coli</p></li>
 +
<li class="green e7"><p>Electrophoresis of [target site2+mRFP]PCR</p></li>
 +
<li class="green e7"><p>Transformation of target site1 in PSB1C3</p></li>
 +
<li class="green e7"><p>Electrophoresis of [target site2+mRFP]PCR&[LacI+J61048+Plac+sRNA2]mini</p></li>
</ul>
</ul>
</div>
</div>
Line 2,951: Line 3,779:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>Digestion: Ptet+B0032[ES] &amp; GliI+KivD[XP]</p></li>
+
<li class="green e2"><p>Electrophoresis of [LacI+J61048+ Plac+sRNA2]dig XP&[Plux+sRNA2]dig XP&[37。C RBS+LuxR+37。C RBS+mGFP]PCR</p></li>
-
<li class="green e2"><p>sample and do GC</p></li>
+
<li class="green e2"><p>Colony PCR of target 1 Cr E.coli&mRFP+J61048 Kr E.coli  and then electrophoresis of these PCR products</p></li>
</ul>
</ul>
</div>
</div>
Line 2,968: Line 3,796:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>ligation : insert Ptet+B0032[ES] &amp; GliI+KivD[XP]/vector pSB1K3[EP]</p></li>
+
<li class="green e2"><p>Colony PCR of target 1 Cr E.coli and then electrophoresis of PCR products</p></li>
-
<li class="green e2"><p>Single colony isolation from G1+G2 LB-C plate, and cultivation in liquid LB-C</p></li>
+
<li class="green e2"><p>Transformation of mRFP+J610148 in PSB1K3(failed)</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
+
<div class="day brn">
<div class="daybar"><p>17</p></div>
<div class="daybar"><p>17</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 2,985: Line 3,815:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>transformation of Ptet+B0032+ GliI+KivD, and cultivation on LB- K plate</p></li>
+
<li class="green e4"><p>Colony PCR of mRFP+J61048 Kr E.coli and then electrophoresis of the PCR product</p></li>
-
<li class="green e2"><p>mini-prep of cultivated G1+G2 E. coli</p></li>
+
<li class="green e4"><p>Colony PCR of target 1 Cr E.coli and then electrophoresis of the PCR product</p></li>
 +
<li class="green e4"><p>Transformation of mRFP+J610148 in PSB1K3(failed)</p></li>
 +
<li class="green e4"><p>Cultivation of mRFP+J61048 Kr E.coli in liquid LB-K tube(failed)</p></li>
</ul>
</ul>
</div>
</div>
-
</div>
+
</div>
-
<div class="day brn">
+
</div>
 +
<!---------------------------------------- week end ---------------------------------------->
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week bigwk">
 +
<div class="day">
<div class="daybar"><p>18</p></div>
<div class="daybar"><p>18</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 3,002: Line 3,840:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [Ptet+B0032+ GliI+KivD]-----OK</p></li>
+
<li class="green e4"><p>Colony PCR of mRFP+J61048  Kr E.coli</p></li>
-
<li class="green e2"><p>DNA sequencing------NOT OK</p></li>
+
<li class="green e4"><p>Digestion:[J61048]XP&[target site1]XP</p></li>
 +
<li class="green e4"><p>Ligation:insert[target site2]ES+[J61048]XP/vector[PSB1C3]EP</p></li>
 +
<li class="green e4"><p>Electrophoresis of [mRFP+J61048]PCR&[target site1]PCR</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
</div>
 
-
<!---------------------------------------- week end ---------------------------------------->
 
-
<!---------------------------------------- week start ---------------------------------------->
 
-
<div class="week">
 
<div class="day">
<div class="day">
<div class="daybar"><p>19</p></div>
<div class="daybar"><p>19</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 3,021: Line 3,862:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e5"><p>Cultivation of mRFP+J61048 Kr E.coli in liquid LB-K tube</p></li>
 +
<li class="green e5"><p>Transformation of target site2+mRFP+J61048 in PSB1C3</p></li>
 +
<li class="green e5"><p>Ligation:insert[target site 1]/vector[PSB1C3]</p></li>
 +
<li class="green e5"><p>Mini-prep of cultivated mRFP+J61048 Kr E.coli</p></li>
 +
<li class="green e5"><p>Digestion:[mRFP+J61048]XP</p></li>
</ul>
</ul>
</div>
</div>
Line 3,029: Line 3,875:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 3,034: Line 3,884:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>Colony PCR of target site2+mRFP+J61048 Cr E.coli</p></li>
 +
<li class="green e4"><p>Transformation of target site1 in PSBC3 Cr E.coli</p></li>
 +
<li class="green e4"><p>Electrophoresis of[mRFP+J61048]dig XP&[Plux+sRNA]digXP</p></li>
 +
<li class="green e4"><p>Mini-prep of cultivated Plux+sRNA2 Kr E.coli Digestion:[Plux+sRNA2]XP</p></li>
</ul>
</ul>
</div>
</div>
Line 3,042: Line 3,896:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 3,047: Line 3,902:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green"><p>Electeophoresis of [mRFP+J61048]dig XP&[ target site2+mRFP+J61048]PCR</p></li>
</ul>
</ul>
</div>
</div>
Line 3,055: Line 3,911:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 3,060: Line 3,921:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e5"><p>Cultivation of target site2+mRFP+J61048 Cr E.coli</p></li>
 +
<li class="green e5"><p>Transformation of target site1 in PSB1C3</p></li>
 +
<li class="green e5"><p>Ligation:insert[mRFP]ES+[J61048]XP/vector[PSB1K3]EP</p></li>
 +
<li class="green e5"><p>Dig:LuxI(BBa_C0061)Transformation of mRFP+J61048 in PSb1K3&LuxI inPSB1A3</p></li>
 +
<li class="green e5"><p>Mini-prep of cultivated target site2+mRFP+J61048 Cr E.coli and then digestion:[target site2+mRFP+J61048]XP</p></li>
</ul>
</ul>
</div>
</div>
Line 3,068: Line 3,934:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
</ul>
</ul>
</div>
</div>
Line 3,073: Line 3,943:
<div class="open">
<div class="open">
<ul>
<ul>
 +
<li class="green e4"><p>Electrophoresis of [target site 2+mRFP+J61048]dig XP&[37。C RBS+LuxR+37。C RBS+GFP]PCR&[Pompc+B0032+LacI+J61048+Plac]PCR&[mRFP+J61048]PCR&[target site1]PCR</p></li>
 +
<li class="green e4"><p>Cultivation of LuxI Ar E.coli in liquid LB-A tube</p></li>
 +
<li class="green e4"><p>Colony PCR of target site1 Cr E.coli&mRFP+J61048Kr E.coli</p></li>
 +
<li class="green e4"><p>Dig:B0034+LuxI(BBa_C0261)Transformation of B0034+LuxI in PSB1A3</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
+
<div class="day brn">
<div class="daybar"><p>24</p></div>
<div class="daybar"><p>24</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 3,087: Line 3,964:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>carbon source test: activation DH5α overnight</p></li>
+
<li class="green e4"><p>Cultivation of 37。C RBS+LuxR+37。C RBS+mGFP Ar E.coli&B0034+LuxI Ar E.coli in liquid LB-A tubes and Pompc+B0032+LacI+J61048+Plac Kr E.coli in liquid LB-K tube</p></li>
 +
<li class="green e4"><p>Digestion:[37。C RBS+mGFP]ES&[37。C RBS+LuxR]XP&[sRNA1]ES&[Pcons]XP</p></li>
 +
<li class="green e4"><p>ligation:insert[target site 1]XP/vector[PSB1C3]XP&insert[mRFP]ES+[B0015]XP/vector[PSB1C3]EP&insert[Pcons]ES+[target site 2+mRFP+J61048]XP/vector[PSB1C3]EP&insert[37。C RBS+mGFP]ES+[37。C RBS+LuxR]XP/vector[PSB1C3]EP&insert[sRNA1]ES+[Pcons]XP/vector[PSB1K3]EP&insert[Pcons]ES+[32。C RBS+mGFP]XP/vector[PSB1C3]EP and then transformation</p></li>
 +
<li class="green e4"><p>mini-prep of cultivated LuxI Ar E.coli&B0034+LuxI Ar E.coli&Pompc+B0032+LacI+J61048+Plac Kr E.coli&37。C RBS+LuxR+37。C RBS+mGFP Ar E.coli</p></li>
</ul>
</ul>
</div>
</div>
-
</div>
+
</div>
-
<div class="day brn">
+
</div>
 +
<!---------------------------------------- week end ---------------------------------------->
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week bigwk">
 +
<div class="day">
<div class="daybar"><p>25</p></div>
<div class="daybar"><p>25</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 3,102: Line 3,990:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>transfer to new medium(1/100), OD0.2 start counting culture time</p></li>
+
<li class="green e5"><p>Digestion:[LuxI]XP&[B0034+LuxI]ES&[ Pompc+B0032+LacI+J61048+Plac]ES&[7。C RBS+LuxR+37。C RBS+mGFP]&[PSB1A3.PSB1C3.PSB1K3]EP&[PSB1C3]ES and XP&[37。C RBS+mGFP]EP and then electrophoresis</p></li>
 +
<li class="green e5"><p>Ligation:insert[B0034+LuxI]ES+[B0015]XP/vector[PSB1C3]EP&[Pcons]ES+[target site 2+mRFP+J61048]XP+[37。C RBS+mGFP]EP</p></li>
 +
<li class="green e5"><p>Transformation of B0034+LuxI+B0015 inPSB1C3&Pcons+target site2+mRFP+J61048&Pcons+32。C RBS+mGFP in PSB1C3</p></li>
 +
<li class="green e5"><p>Colony PCR of sRNA1+Pcons Kr E.coli&target site1 Cr E.coli&mRFP+B0015 Cr E.coli&37。C RBS+mGFP+37。C RBS+LuxR Cr E.coli</p></li>
 +
<li class="green e5"><p>Electrophoresis of [target site1]PCR&[mRFP+B0015]PCR&[37。C RBS+mGFP+37。C RBS+LuxR]PCR+[sRNA1+Pcons]PCR</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
</div>
 
-
<!---------------------------------------- week end ---------------------------------------->
 
-
<!---------------------------------------- week start ---------------------------------------->
 
-
<div class="week mweek">
 
<div class="day">
<div class="day">
<div class="daybar"><p>26</p></div>
<div class="daybar"><p>26</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 3,121: Line 4,012:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>culturing for 72hours, inject feeding solution per 24hours</p></li>
+
<li class="green e4"><p>Mini-prep of cultivate target site1Cr E.coli&mRFP+B0015 Cr E.coli&37。C RBS+mGFP+37。C RBS+LuxR Cr E.coli&sRNA+Pcons Kr E.coli </p></li>
 +
<li class="green e4"><p>Digestion:[target site 1]ES&[ mRFP+B0015]XP&[37。C RBS+mGFP+37。C RBS+LuxR]ES&[ sRNA+Pcons]ES&[37。C　RBS+LuxR+37。C RBS+mGFP]ES</p></li>
 +
<li class="green e4"><p>Colony PCR of B0034+LuxI+B0015 Cr E.coli&Pcons+target site 2+mRFP+J61048 Kr E.coli</p></li>
 +
<li class="green e4"><p>Electrophoresis of those digestion and PCR products made today</p></li>
</ul>
</ul>
</div>
</div>
Line 3,130: Line 4,024:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
</ul>
</ul>
Line 3,136: Line 4,034:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green"><p>culturing for 72hours, inject feeding solution per 24hours</p></li>
+
<li class="green e5"><p>Mini-prep of cultivated Pcons+target site2+mRFP+J61048 K r E.coli</p></li>
 +
<li class="green e5"><p>Digestion:[ Pcons+target site2+mRFP+J61048]ES&[Pompc+B0032+LacI+J61048+Plac]XP</p></li>
 +
<li class="green e5"><p>Ligation insert[target site 1]ES+[mRFP+B0015]XP/vector[PSB1A3]EP&insert[Pcons+target site2+mRFP+J61048]ES+[Pcons+B0032+LacI+J61048+Plac+sRNA2]XP/vector[PSB1A3]EP&insert[Pcons]ES+[37。C RBS+mGFP+37。CRBS+LuxR]XP/vector[PSB1K3]EP&insert[Pcons]ES+[37。C RBS+LuxR+37。C RBS+mGFP]XP/vector[PSB1K3]EP&insert[Pcons+target site 2+mRFP+J61048]ES+[Plux+sRNA2]XP/vector[PSB1A3]EP&insert[Pompc+B0032+LacI+J61048+Plac]ES+[37。C RBS+mGFP+37。C RBS+LuxR]XP/vector[PSB1A3]EP&insert[Pompc+B0032+LacI+J61048+Plac]ES+[37。CＲＢＳ＋LuxR+37。C RBS+mGFP]XP/vector[PSB1C3]EP&insert[sRNA1(With Plux)]ES+[Pompc+B0032+LacI+J61048+Plac]XP/vector[PSB1A3]EP&insert[Pompc+B0032+LacI+J61048+Plac]ES+[target site2+mRFP]XP/vector[PSB1A3]&insert[Pcons+target site 2+mRFP+J61048]ES+[Pcons+B0032+LacI+J61048+Plac+sRNA2]XP/vector[mGFP]EP&insert[Pcons+target site2+mRFP+J61048]ES+[Plux+sRNA]XP/vector[mGFP]EP </p></li>
 +
<li class="green e5"><p>Transformation of these ligation product and B0034+LuxI+B0015 in PSB1C3</p></li>
 +
<li class="green e5"><p>Electrophoresis of [Pcons+tar get site 2+mRFP+J61048]PCR&[PSB1K3]dig EP&[Pcons+target site2+mRFP+J61048]dig ES&[Pompc+B0032+LacI+J61048+Plac]XP&[37CRBS+mGFP+37C </p></li>
</ul>
</ul>
</div>
</div>
Line 3,152: Line 4,054:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>transformation of DNA program, and cultivation on LB- A plate</p></li>
+
<li class="green e2"><p>Cultivation of B0034+LuxI+B0015 E.coli on LB-C plate&Pcons+37。C RBS+mGFP+37。C RBS+LuxR E.coli on LB-K plate&Pcons+37。C RBS+LuxR+37。C RBS+mGFP E.coli on LB-K plate&sRNA1(with Plux)+Pompc+B0032+LacI+J61048+Plac on LB-A plate</p></li>
-
<li class="green e2"><p>culturing for 72hours, inject feeding solution per 24hours</p></li>
+
<li class="green e2"><p>Colony PCR of target site 1+mRFP+B0015 Ar& Pcons+target site2+mRFP +J61048+Pcons+B0032+LacI+J61048+Plac+sRNA2 Ar & Pcons+37。C RBS+mGFP+37。C RBS+LuxR Kr&Pcons+target site 2+mRFP+J61048+Plac+sRNA2 Ar&Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR Ar&PomPC +B0032+LacI+J61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP Cr &sRNA2(with Plac)+Pompc+B0032+LacI+J61048+Plac Ar& Pompc+B0032+LacI+J61048+Plac+target site 2+mRFP+J61048 Ar E.coli and then electrophoresis of these PCR products</p></li>
</ul>
</ul>
</div>
</div>
Line 3,162: Line 4,064:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 3,170: Line 4,073:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>ligation : insert Ptet+B0032[ES] &amp; GliI+KivD[XP]/vector pSB1K3[EP]</p></li>
+
<li class="green e4"><p>Mini-prep of cultivated target site 1+mRFP+B0015 Ar &Pcons+target site 2+mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA 2 Ar&Pcons+37。C RBS＋mGFP+37。C RBS+LuxR Kr & Pompc+B0032+LacI+J61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP Cr & Pcons+target site 2+mRFP+J61048+Plux+sRNA2 Kr &Pcons+37。C RBS+LuxR+37。C RBS+mGFP Kr E.coli</p></li>
-
<li class="green e3"><p>Single colony isolation from DNA program LB-C plate, and cultivation in liquid LB-C</p></li>
+
<li class="green e4"><p>Digestion:[ target site 1+mRFP+B0015]XP&[Pcons+target site 2+mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA 2]XP&[ Pcons+37。C＋mGFP+37。C RBS+LuxR ]ES&[ Pompc+B0032+LacI+J61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP]ES and then electrophoresis of these digestion products</p></li>
-
<li class="green e3"><p>sample &amp; do GC</p></li>
+
<li class="green e4"><p>Colony PCR of Pcons+target site mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA2 Ar &Pcons+target site2+mRFP+J61048+Plac+sRNA2 Ar &Pompc+B0032+LacI+J61048+Plac+37。 C RBS+mGFP+37。C RBS+LuxR Ar &Pompc+B0032+LacI+J61048+Plac+target site2+mRFP+J61048 Ar &sRNA1(with Plac)+Pompc+B0032+LacI+J61048+Plac Ar &Pcons+37。C RBS+LuxR+37。C RBS+mGFP Kr &B0034+LacI+B0015 Cr E.coli and then electrophoresis of these PCR products</p></li>
 +
<li class="green e4"><p>Ligation:insert[Pcons]ES+[37。 C RBS+mGFP]XP/vector[PSB1C3]EP&insert[37。C RBS]ES+[LacI+J61048+Plac+sRNA2]XP/vector[PSB1K3]EP&insert[B0034+LuxI]ES+[J61048]</p></li>
</ul>
</ul>
</div>
</div>
Line 3,181: Line 4,085:
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 3,189: Line 4,094:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e3"><p>transformation of Ptet+B0032+ GliI+KivD, and cultivation on LB- K plate</p></li>
+
<li class="green e4"><p>Cultivation of target site1+mRFP+B0015 Ar and Pcons+target site2+mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA2 Ar E.coli in liquid LB-A tubes &Pcons+37。C RBS+mGFP+37。C RBS+LuxR Kr E.coli in liquid LB-K tube &Pompc+B0032+LacI+J+61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP Cr E.coli in liquid LB-C tube &Plux+sRNA 2 in liquid LB tube </p></li>
-
<li class="green e3"><p>mini-prep of cultivated DNA program E. coli</p></li>
+
<li class="green e4"><p>Mini-prep of those cultivated E.coli which was cultivated this morning</p></li>
-
<li class="green e3"><p>Do GC</p></li>
+
<li class="green e4"><p>Electrophoresis of the mini-prep products made today and [Pcons+target site 2+mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA 2]dig ES &[ Pcons+target site 2+mRFP+J61048+Plux+sRNA2]dig ES&[cons+37。C RBS+LuxR+37。C RBS+mGFP]XP</p></li>
 +
<li class="green e4"><p>digestion:[B0034]XP&[target site 2]XP colony PCR of Pcons+37。C RBS Cr E.coli&37。C RBS+LacI+J61048+Plac+sRNA2 Kr E.coli&B0034+LacI+J61048 Kr E.coli&B0032+LacI Kr E.coli and then electrophoresis of these PCR products</p></li>
</ul>
</ul>
</div>
</div>
</div>
</div>
-
<div class="day">
+
<div class="day brn">
<div class="daybar"><p>31</p></div>
<div class="daybar"><p>31</p></div>
<div class="dots">
<div class="dots">
<ul>
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
<li class="caldot green"></li>
Line 3,207: Line 4,116:
<div class="open">
<div class="open">
<ul>
<ul>
-
<li class="green e2"><p>PCR of insert fragment [Ptet+B0032+ GliI+KivD] OK</p></li>
+
<li class="green e5"><p>Ligation:insert[Pcons]ES+[target site 2]XP/vector[PSB1K3]EP&insert[Pcons]ES+[B0034]XP/vector[PSB1K3]EP</p></li>
-
<li class="green e2"><p>DNA sequencing NOT OK</p></li>
+
<li class="green e5"><p>Transformation of Pcons+target site 2 in PSB1K3&Pcons+B0034 in PSB1K3 & BBa_K516030(mRFP protein generator) in PSB1C3&BBa_K516132(Constitutive promoter [J23101] with mRFP, RBS B0032) in PSB1C3&Pcons+LacI+Plac+sRNA2 in PSB1K3</p></li>
 +
<li class="green e5"><p>Mini-prep of cultivated Pcons+37。C RBS+mGFP Cr E.coli&37。C RBS+LacI+J61048+Plac+sRNA2 Cr E.coli&B0032+LuxI Kr E.coli </p></li>
 +
<li class="green e5"><p>digestion:[ Pcons+37。C RBS+mGFP]ES&[37。C RBS+LacI+J61048+Plac+sRNA2]XP&[ B0032+LuxI]ES &[Pcons+37。C RBS+mGFP+37。C RBS+LuxR]ES&[Pompc+B0032+LacI+J61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP]ES</p></li>
 +
<li class="green e5"><p>cultivation of Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR Ar &Pompc+B0032+LacI+J61048+Plac+target site 2+mRFP=J61048 Ar E.coli on LB-A plates</p></li>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
</div>
 +
<!---------------------------------------- week end ---------------------------------------->
 +
</div>
 +
</div>
 +
</div>
 +
<!---------------------------------------- cal ends ---------------------------------------->
 +
</div>
 +
</div>
 +
<div class="li"><div class="card">
 +
<h3>September 2013</h3>
 +
<!---------------------------------------- cal starts ---------------------------------------->
 +
<div class="calendar">
 +
<div class="calcontainer">
 +
<div class="daysweek">
 +
<div class="dayweek"><p>Sunday</p></div>
 +
<div class="dayweek"><p>Monday</p></div>
 +
<div class="dayweek"><p>Tuesday</p></div>
 +
<div class="dayweek"><p>Wednesday</p></div>
 +
<div class="dayweek"><p>Thursday</p></div>
 +
<div class="dayweek"><p>Friday</p></div>
 +
<div class="dayweek brn"><p>Saturday</p></div>
 +
</div>
 +
<div class="daysmonth">
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week bigwk">
 +
<div class="day">
 +
<div class="daybar"><p>1</p></div>
 +
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
<li class="green"><p>Ligation:insert[Pcons+37。C RBS+mGFP]ES+[37。C RBS+LacI+J61048+Plac+sRNA2]XP/vector[PSB1A3]EP&inert[Pcons+37。C RBS+mGFP]ES+[37。C RBS+LuxR]XP/vector[PSB1A3]EP</p></li>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>2</p></div>
 +
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
<li class="green"><p>Mini-prep of cultivated Pcons+37。C RBS+mGFP+37。C RBS+LacI+J61048+Plac+sRNA2 Ar E.coli Colony  PCR of Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR Ar &sRNA(with Plux)+Pompc+B0032+LacI+J61048+Plac Ar &Pompc+B0032+LacI+J61048+Plac+target site2+mRFP+J61048 Ar &Pcons+37。C RBS+mGFP+37。C RBS+LuxR Ar E.coli Electrophoresis of PCR products today</p></li>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>3</p></div>
 +
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
<li class="green e3"><p>Mini-prep of cultivated Pcons+37。C RBS+mGFP+37。C RBS+LacI+J61048+Plac+sRNA2 Ar E.coli Colony  PCR of Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR Ar &sRNA(with Plux)+Pompc+B0032+LacI+J61048+Plac Ar &Pompc+B0032+LacI+J61048+Plac+target site2+mRFP+J61048 Ar &Pcons+37。C RBS+mGFP+37。C RBS+LuxR Ar E.coli Electrophoresis of PCR products today</p></li>
 +
<li class="green e3"><p>Digestion:[ Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR]ES&[ sRNA1(with Plux)+Pompc+B0032+LacI+J61048+Plac]ES&[ Pompc+B0032+LacI+J61048+Plac+target site2+mRFP+J61048]ES&[Pcons+37。C RBS+mGFP+37。C RBS+Lux]ES&[BBa_K516030(Constitutive promoter [J23101] with mRFP, RBS B0030) ]XP and then electrophoresis</p></li>
 +
<li class="green e3"><p>Ligation:insert[Pcons]ES+[target site 2]XP/vector[PSB1K3]&insert[Pcons]ES+[B0034]XP/vector[PSB1K3]EP&insert[B0034+LuxI]ES+[B0015]XP/vector[PSB1C3]EP</p></li>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>4</p></div>
 +
<div class="dots">
 +
<ul>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
<li class="caldot green"></li>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
<li class="green e4"><p>Colony PCR of Pcons+B0034 Kr&Pcons+target site Kr&Pcons+K516030 Kr&B0034+LacI+B0015 Cr E.coli and then electrophoresis of these PCR products</p></li>
 +
<li class="green e4"><p>Ligation: insert[Pcons+B0032]ES+[LacI+J61048+Plac+sRNA2]XP/vector[PSB1K3]&insert[37。C RBS]ES+[LacI+J61048+Plac+sRNA2]XP/vector[PSB1K3]EP&insert[Pompc+B0032+LacI+J61048+Plac]ES+[37。C RBS+mGFP]XP/vector[PSB1C3]EPBackbone changing:[37。C RBS+mGFP]XP&[37。C RBS+LuxR]XP&[37。C RBS+LuxR+37。C RBS+mGFP]XP&[Pcons+B0030+PcyA+B0032+LacI+B0030]ES&[Pcons+target site2+mRFP+J61048]ES /vector[PSB1C3]ES and [sRNA+target site2]EP/vector[PSB1C3]EP</p></li>
 +
<li class="green e4"><p>Digestion:[sRNA+target site2]EP
 +
Transformation of Pcons+B0032+LacI+J61048+Plac+sRNA2 in PSB1K3&37。C RBS+LacI+J61048+Plac+sRNA2 in PSB1K3&Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP in PSB1C3&37。C RBS+mGFP in PSB1C3&37。C RBS+LuxR in PSB1C3&37。C RBS+LuxR+37。C RBS+mGFP in PSB1C3&Pcons+B0034+PcyA+B0032+LacI+B0030 in PSB1C3&Pcons+target site2+mRFP+J61048 in PSB1C3&sRNA+target site 2 in PSB1C3</p></li>
 +
<li class="green e4"><p>Mini-prep of cultivated Pcons+B0034 Kr E.coli&Pcons+target site2 Kr E.coli&Pcons+K516030 Kr E.coli and then digestion:[ Pcons+B0034]ES and E&[ Pcons+target site2]ES and E&[Pcons+K516030]ES</p></li>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>5</p></div>
 +
<div class="dots">
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<ul>
 +
</ul>
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</div>
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 +
<div class="open">
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<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>6</p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
</ul>
</ul>
</div>
</div>
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</div>
</div>
<div class="day brn">
<div class="day brn">
-
<div class="daybar"><p></p></div>
+
<div class="daybar"><p>7</p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
</div>
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week">
 +
<div class="day">
 +
<div class="daybar"><p>8</p></div>
 +
<div class="dots">
 +
</div>
 +
 
 +
<div class="open">
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>9</p></div>
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<div class="dots">
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<ul>
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</div>
 +
 
 +
<div class="open">
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<ul>
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</ul>
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</div>
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 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>10</p></div>
 +
<div class="dots">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>11</p></div>
 +
<div class="dots">
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<ul>
 +
</ul>
 +
</div>
 +
 
 +
<div class="open">
 +
<ul>
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</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>12</p></div>
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<div class="dots">
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<ul>
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</ul>
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</div>
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 +
<div class="open">
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<ul>
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</ul>
 +
</div>
 +
 
 +
</div>
 +
<div class="day">
 +
<div class="daybar"><p>13</p></div>
 +
<div class="dots">
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<ul>
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</ul>
 +
</div>
 +
 
 +
<div class="open">
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</div>
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 +
</div>
 +
<div class="day brn">
 +
<div class="daybar"><p>14</p></div>
 +
<div class="dots">
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<ul>
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</div>
 +
 
 +
<div class="open">
 +
<ul>
 +
</ul>
 +
</div>
 +
 
 +
</div>
 +
</div>
 +
<!---------------------------------------- week start ---------------------------------------->
 +
<div class="week mweek">
 +
<div class="day">
 +
<div class="daybar"><p>15</p></div>
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<div class="dots">
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</div>
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 +
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<div class="day">
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<div class="daybar"><p>16</p></div>
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<div class="day">
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<div class="daybar"><p>17</p></div>
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<div class="day">
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<div class="daybar"><p>18</p></div>
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<div class="day">
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<div class="daybar"><p>19</p></div>
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<div class="daybar"><p>20</p></div>
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<div class="daybar"><p>21</p></div>
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<div id="footer-wrapper">
<div id="footer-wrapper">
   <div id="footer"> <div id="footer-text">
   <div id="footer"> <div id="footer-text">
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     <p>Copyright © 2013 NCTU_Formosa</p>
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     <p>2013 NCTU_Formosa</p>
     <p class="author">Website designed by Calvin Hue.</p>
     <p class="author">Website designed by Calvin Hue.</p>
 +
    <p>Cover image credit: <a href="http://projectheshima.wordpress.com/" target="_blank">Project Heshima</a></p>
     </div> </div>
     </div> </div>
</div>
</div>

Latest revision as of 03:58, 28 September 2013

Notes

The experimental log that records down the purpose, method, result and date of each experiment that we have conducted.

About the notes

Our notes are the abbreviations of the experimental process, including titles, genes we transformed, plasmids we used, and resistances on the plasmids. We categorized these notes by date. Moreover, team members followed the iGEM protocol in each experimental step in order to get better experimental efficiency and the results we had expected.

Material and Methods

Minipreps of Plasmid DNA

We use Plasmid miniPREP kit (Genedirex Inc.) to obtain plasmid DNA for analysis, sequencing and cloning. Single colony is picked from LB agar plate and inoculated into LB medium with suitable antibiotics. Bacteria cells are harvest by centrifuge after incubation for 12 – 16 hours. Afterward, alkaline lysis is performed to release inside components, and DNA is purified by silicone resin-based column. The quality and amount of plasmid DNA is estimated by agarose gel electrophoresis stained by SyBr Safe (Invitrogen).

BioBrick assembly

To digest the desired DNA, appropriate restriction enzymes and reaction buffer (all perched from New England Biolabs) is used in 20 μL reaction volume. Upstream BioBrick is digested with EcoRI-HF and SpeI in CutSmart buffer. Downstream BioBrick and backbone part are digested with XbaI and PstI, EcoRI-HF and PstI respectively in NEBuffer 2. Reactions take place in 37 ℃ water bath for 2 hours or overnight.

Ligation is performed using BioBrick Assembly Kit Ligation Protocol. All the reagent and enzymes are perched from New England Biolabs). Reactions take place in 37 ℃ water bath for 2 hours or overnight in 16 ℃ incubator.

Transformation

DH5α E. coli competent cells (ECOSTM 101, Yeastern Biotech Co., Ltd.) were used for the propagation of plasmid DNA. Standard transformation protocol is used: First place 2 μL plasmid or 10μL product into 1.5 mL mini centrifuge tube and chilled on ice. Competent cells are melt on ice before transformation. After adding 33 – 50 μL competent cells, heat shock at 42 ℃ for 1 minutes. Place cells into LB medium to recover then plating on LB agar plate and incubate for 16 – 18 hours.

PCR reaction

Taq DNA Polymerase 2x Master Mix Red (Ampliqon) and VF2 and VR primers (BBa_G00100 and BBa_G00101 respectively) are used for colony PCR. Single colony is picked and place into 10 μL PCR reagent, and followed by PCR with condition described below:

  • Predenature: 95 ℃，10 min
  • Denature, annealing and extension: [98 ℃, 10 s; 55 ℃, 30 s; 72 ℃, 1 m/kb + 30 s] × 25 cycles
  • Hold: 4 ℃

Reagent and Medium

Lysogeny broth (LB): 10 g/L tryptone, 5 g/L yeast extract, 10 g/L NaCl with with suitable antibiotics.

LB agar plate: LB medium with 1.5% (w/v) agar.

50X TAE buffer: tris base 242 g/L, glacial acetic acid 57.1 mL/L, 0.05 M EDTA, pH 8.0 Antibiotics working concentration:

  • Kanamycin  : 25 μg/mL
  • Ampicillin  : 100 μg/mL
  • Chloramphenicol: 20 μg/mL


March 2013

Sunday

Monday

Tuesday

Wednesday

Thursday

Friday

Saturday

24

25

26

27

28

29

  • Transformation of ho1(Kr), Cph8(Cr), 32℃ RBS(Ar) and cultivation on three LB plates.

  • transformation of PompC and cultivation on LB-A plate

30

  • Single colony isolation from three plates and cultivation them in liquid LB

  • Single colony isolation from PompC LB-C plate and cultivation in liquid LB-A

  • mini-prep of cultivated PompC E.coli

31

  • Transformation of RBS B0030+PSB1A3&B0034+PSB1A3 and cultivation on LB-A plate.

  • Mini-prep for cultivated ho1(Kr), Cph8(Cr), 32℃ RBS(Ar) PCR of ho1(Kr), Cph8(Cr), 32℃ RBS(Ar).

  • Electrophoresis of ho1(Kr), Cph8(Cr), 32℃ RBS(Ar) ──NOT OK.

  • PCR of insert fragment [PompC+psB1A2]

April 2013

Sunday

Monday

Tuesday

Wednesday

Thursday

Friday

Saturday

1

  • Cultivation of B0030&B0034 Ar E.coli in liquid LB-A tubes.

2

  • Mini-prep of cultivated B0030&B0034 E.coli and then digestion: [B0030]XP&[B0034]XP.

3

4

5

6

  • Electrophoresis of [B0030]XP&[B0034]XP Not OK.

  • PCR of ho1 to check ho1 and transform to test the resistance.

7

  • Electrophoresis of [B0030]mini&dig XP and [B0034]mini&dig XP OK

  • Cultivation of B0030&B0034 Ar E.coli in liquid LB-A tubes.

  • Cultivation of ho1 in liquid LB-K and LB-K plate

  • Cultivation of Cph8+RBS in liquid LB-C.

    • 8

    • Mini-prep of cultivated B0030&B0034 Ar E.coli. Mini-prep of ho1 & Cph8+RBS.

    9

    • Electrophoresis of mini ho1&Cph8+RBS.

    10

    11

    • PCR of mini ho1&Cph8+RBS

    • Electrophoresis of ho1&Cph8+RBS(After PCR)

    • Digestion: [ho1]EP&[Cph8+RBS]EP.

    12

    • Transformation of Plac&Ptet& pcyA but there is no colony appearing in pcyA plate.

    • Electrophoresis of digested ho1&Cph8+RBS (NOT OK).

    13

    14

    • Cultivation of Ptet&Plac E.coli in LB tubes.

    • Transformation: LuxR,B0015,J61048 and cultivation on LB-A plate

    15

    • Mini-prep of cultivated Ptet&Plac E.coli

    • Digestion:[Ptet]ES&[Plac]ES PCR of insert fragment pcyA

    • Electrophoresis of [Ptet]dig ES&[pcyA]PCR&[Plac]dig ES.

    • Single colony isolation from J61048,B0015,LuxR plate and cultivation in liquid LB-A mini-prep of cultivated LuxR,J61048,B0015

    16

    • PCR of mini Cph8+RBS

    • Electrophoresis of Cph8+RBS(After PCR)──NOT OK.

    • digestion: [J61048]EP,[B0015]EP,[LuxR]EP

    • Electrophoresis of ter. mini [J61048] dig E/EP and t er. mini [B0015] dig E/EP and luxR mini dig E/EP

    17

    • PCR of mini Cph8+RBS Electrophoresis of Cph8+RBS(After PCR)──NOT OK.

    • Electrophoresis of the products of digestion of ter. [J61048], ter. [B0015] , luxR

    • transformation of tetR plasmid and cultivation on LB –A plate 3. Digestion: ter. [J61048], ter. [B0015] dig EP

    18

    19

    • Cultivation of pcyA Kr E.coli on LB-K plates

    • mini-prep of cultivated tetR E.coli

    • digestion: tetR+pSB1A2 dig EP

    • electrophoresis of tetR dig EP,ter.[J61048 ] dig EP and ter.[B0015] dig EP

    20

    • Cultivation of pcyA Kr E.coli in liquid LB-K tubes.

    • PCR of tetR,ter.[J61048] and ter.[B0015 ]mini

    • electrophoresis of PCR products of tetR mini,J61048 mini and B0015 mini

    21

    • Single colony isolation of pcyA Kr E.coli

    • Mini-prep of cultivated pcyA Kr E.coli

    • Digestion:[pcyA]ES

    • Electrophoresis of [pcyA]mini&dig ES OK

    • PCR of tetR,ter.[J61048] and ter.[B0015 ]mini

    • electrophoresis of PCR products of tetR mini,J61048 mini and B0015 mini

    • electrophoresis of the digestion products of tetR dig EP and B0015 dig EP

    • aliquot every 20ul of PompC,J61048 and LuxR

    22

    23

    24

    25

    26

    27

    28

    • Transformation of PompC mini and cultivation on LB-A plate

    29

    • transformation of PompC , Ar mini and cultivation on LB-A plate

    • single colony isolation from PompC , Ar

    30

    • Transformation of 37ﾟC RBS and ho1.

    • Mini-prep of cultivated PompC , Ar E.coli

    May 2013

    Sunday

    Monday

    Tuesday

    Wednesday

    Thursday

    Friday

    Saturday

    1

    • Cultivation of 37ﾟC RBS and ho1

    • Digestion: [pSB1C3] EP&[B0030]ES&{pcyA]XP

    • Transformation of pSB1A3 &pSB1K3

    • Transformation of mGFP[pSB1A2] and cultivation on LB-A plate.

    2

    • Cultivation of PompC&B0034&LacI&Plac&B0030&TetR&J61048&B0015 Ar E.coli in liquid LB-A tubes.

    • Cultivation of pSB1A3&pSB1K3 with liquid LB

    • Mini-prep of 37ﾟC RBS and ho1──Fail

    • Ligation: [pSB1C3] EP&[B0030]ES& {pcyA]XP

    • Transformation of B0030

    • Cultivation of 37ﾟC RBS and ho1 with liquid LB

    • Transformation of RBS+pcyA+psB1C3.

    • Transformation of mGFP[pSB1A2] and cultivation on LB-A plate

    3

    • Mini-prep of cultivated PompC &B0034&LacI&Plac&B0030&TetR&J61048&B0015,and then digestion:[PompC ]ES&[B0034]XP&[LacI]ES&[Plac]XP&[B0030]ES&[TetR]ES&[TetR]XP&[J61048]XP&[J61048]ES&[B0015]XP.

    • Mini-prep of ho1,pSB1A3, pSB1K3 and 37ﾟC RBS

    • Digestion: [B0030+pcyA]XP

    • Cultivation of B0030 with liquid LB-C

    • Check RBS+pcyA+psB1C3(PCR & Electrophoresis)──FAIL

    • Digestion: [Pcons]ES& [ho1]XP.

    • Single colony isolation from 37。C RBS Ar,pSB1A3,pSB1C3 and pSB1K3

    • electrophoresis of mGFP dig E

    4

    • Electrophoresis of [PompC]ES&[B0030]ES&[LacI]ES&[TetR]ES&[J61048]XP&[TetR]ES&[B0015]XP OK

    • Ligation: insert [PompC]ES+[B0030]XP&[LacI]ES+[J61048]XP&[J61048]ES+[Plac]XP&[B0030]ES+[TetR]XP&[TetR]ES+[B0015]XP/vector[PSB1C3]EP

    • Transformation of B0030+ho1.

    • Mini-prep of cultivated 37。C RBS Ar pSB1A3,37。C RBS+mGFP, Kr and pSB1K3 E.coli

    • digestion:37。C RBS dig ES,mGFP dig XP,LuxR dig XP and pSB1K3 dig EP

    • electrophoresis of digestion products of LuxR,37。C RBS, mGFP and pSB1K3

    • ligation:37。C RBS+luxR+pSB1K3

    • transformation of 37。C RBS+mGFP, Kr->pSB1K3 and cultivation on LB-K plate

    5

    • Electrophoresis of digested RBS+pcyA──OK

    • Ligation: RBS+ho1

    • Cultivation of J61048 with liquid LB-C

    • Digestion: [pSB1A3]EP &[pSB1K3]EP

    • Transformation of ligated RBS+ho1.

    • Ligation:37。C RBS+luxR+pSB1K3 and 37。C RBS+mGFP+pSB1K3

    • transformation of 37。C RBS+luxR, Kr and 37。C RBS+mGFP ,Kr and cultivation on LB-K plate

    6

    • Mini-prep of J61048

    • Ligation: RBS+pcyA+Pcons&pSB1K3

    • Electrophoresis of J61048.

    • Digestion: [J61048]XP

    • Transformation of RBS+pcyA+Pcons.

    • Check LB-A&C&K plates.

    7

    • Mini-prep of J61048 -Ligation: RBS+pcyA+Pcons&pSB1K3

    • Electrophoresis of J61048.

    • Digestion: [J61048]XP

    • Transformation of RBS+pcyA+Pcons.-Check LB-A&C&K plates.

    8

    • Ligation:insert[PompC]ES+[B0034]XP&[LacI]ES+[J61048]XP&[B0030]ES+[TetR]XP&[TetR]ES+[B0015]XP/vector[PSB1C3]E

    • PCR of 37。RBS+luxR, Kr and 37。C RBS+mGFP ,Kr

    • electrophoresis of 37。RBS+luxR ,Kr and 37。C RBS+mGFP ,Kr

    9

    • Transformation and cultivation of PompC+B0034&LacI+J61048&B0030+TetR&TetR+B0015(PSB1C3) on LB-C plates.

    • PCR of 37。RBS+luxR, Kr and 37。C RBS+mGFP ,Kr

    • electrophoresis of 37。RBS+luxR, Kr and 37。C RBS+mGFP ,Kr

    • single colony isolation from37。RBS+luxR, Kr and 37。C RBS+mGFP ,Kr

    10

    • Cultivation of Pcons+RBS+pcyA and B0030 with liquid LB.

    • Mini-prep of cultivated 37。C RBS+luxR, Kr E.coli and 37。C RBS+mGFP ,Kr E.coli

    • digestion: 37。RBS+luxR, dig ES and 37。C RBS+mGFP dig ES

    11

    • Mini-prep of Pcons+RBS+pcyA.

    • Electrophoresis of Pcons+RBS+pcyA──NOT OK.

    • Electrophoresis of 37。C RBS+mGFP, Kr mini, 37。C RBS+mGFP dig ES, 37。C RBS+luxR, Kr mini and 37。C RBS+luxR dig ES

    12

    • Electrophoresis of 37。C+luxR mini, dig ES and 37。C RBS+mGFP mini,dig ES

    13

    • Cultivation of TetR+B0015 Cr E.coli in liquid LB-C tube

    • Single colony isolation of backbone PSB1K3 E.coli

    • Ligation:insert[B0030]ES+[TetR]XP&[TetR]ES+[B0015]XP/vector[PSB1C3]EP.

    • Electrophoresis of Pcons+RBS+pcyA──NOT OK

    • Transformation of B0032&B0034.

    14

    • Ligation:insert[PompC]ES+[B0034]XP&[LacI]ES+[J61048]XP/vector[PSB1C3]EP.

    • Cultivation of Pcons+RBS+pcyA & B0032&B0034 with liquid LB.

    15

    • Mini-prep of cultivated PSB1K3 E.coli

    • Cultivation of B0015+TetR+PSB1C3 and TetR+B0015+PSB1C3 in liquid LB-C plates and Plac+PSB1A3 in liquid LB-A plate. Mini-prep of Pcons+RBS+pcyA&B0032&B0034.

    16

    • Ligation:insert[PompC]ES+[B0034]XP&[LacI]ES+[J61048]XP/vector[PSB1K3]EP

    • Transformation and cultivation of PompC+B0034&LacI+J61048 (PSB1K3) on LB-K plates.

    17

    • cultivation of PompC+B0034&LacI+J61048 Kr E.coli on liquid LB-K tubes.

    • single colony isolation of PompC+B0034&LacI+J61048 Kr E.coli.

    • Electrophoresis of Pcons+RBS+pcyA(NOT OK) &B0032&B0034

    • Cultivation of Pcons+RBS+pcyA.Digestion: [B0032]ES &[B0023]ES.

    18

    • Cultivation of PompC+B0034&LacI+J61048 Kr E.coli from single colony isolation plate made yesterday

    • Min-prep of cultivated [B0030+TetR]Cr&[ Plac]Ar&[PompC+B0034]Kr&[LacI+J61048]Kr E.coli and then digestion:[PompC+B0034]ES &[LacI+J61048]XP. Mini-prep Pcons+RBS+pcyA

    • Electrophoresis of Pcons+RBS+pcyA& digested B0032&B0034──NOT OK

    • Digestion: [B0032]ES &[B0034]ES &[Pcons+RBS+pcyA]E.

    19

    • Electrophoresis of [LacI+J61048]mini & dig XP and [PompC+B0034]mini & dig ES and [PSB1K3]mini & dig EP.

    • Electrophoresis of digested B0032&B0034&Pcons+RBS+pcyA──OK

    • Transformation of pSB1C3

    20

    • Digestion:[Plac]P.

    • Ligation: Pcons+RBS+pcyA&pSB1K3, B0032+ho1&pSB1C3, B0034+ho1&pSB1C1

    • Transformation of Pcons+RBS+pcyA&pSB1K3, B0032+ho1&pSB1C3, B0034+ho1&pSB1C1

    • Single colony isolation from pSB1C3

    21

    • Single colony isolation of TetR+B0015 Cr Ecoli

    • Electrophoresis of [PompC+B0034]mini&dig ES and[Plac]mini&dig P.

    • Cultivation of Pcons+RBS+pcyA &B0032+ho1 with liquid LB

    • Mini-prep of cultivated pSB1C3 E.coli

    22

    • Digestion:[Plac]XP

    • Electrophoresis of [PompC+B0034]mini&dig ES and[Plac]mini&dig XP

    • Ligation: insert[TetR]ES+[B]XP/vector[PSBC3]EP

    • Mini-prep of Pcons+RBS+pcyA

    • Mini-prep of cultivated 37。RBS+mGFP E.coli

    • digestion:37。C RBS+mGFP dig ES and mini

    • electrophoresis of 37。C RBS+mGFP mini and 37。C RBS+mGFP dig ES.

    23

    • Single colony isolation of TetR+B0015 transformation of TetR +B0015 +PSB1C3 on LB-C plates

    • Ligation +transformation on: insert[TetR]ES+[B0015]XP&[PompC]ES+[B0034]/vector[PSB1K3]EP on LB-K plates

    24

    • Single colony isolation of PompC+B0034+PSB1K3 E.coli

    • Electrophoresis of Pcons+RBS+pcyA.

    • -Ligation: B0032+ho1&B0034+ho1.

    • Digestion; [Pcons+RBS+pcyA]ES &[ho1]XP&[pSB1C3]EP.

    • Electrophoresis of 37。 RBS+mGFP dig ES

    25

    • Ligation and transformation: insert [PompC]ES+[B0034]XP/vector[PSB1K3]EP on LB-K plates

    • Single colony isolation of PompC+B0034+PSB1K3 E.coli again

    • Mini-prep of cultivated TetR+B0015 Cr E.coli and digestion:[TetR+B0015]P

    • Electrophoresis of :[TetR+B0015]P

    • Digestion: [TetR+B0015]XP

    • Cultivation of PompC+B0034 in liquid LB-K tubes.

    • Mini-prep of B0032+ho1

    • Cultivation of B0034+ho1 with liquid LB-A

    26

    • Cultivation of PompC+B0034Kr E.coli in liquid LB-K tubes again

    • Mini-prep of cultivate PompC+B0034 Kr E.coli

    • Electrophoresis of [TetR+B0015] dig XP not OK

    • Mini-prep of B0034+ho1 Electrophoresis of Pcons+RBS+pcyA & B0032+ho1 & B0034+ho1.

    27

    • Digestion:[TetR+B0015]XP

    • Electrophoresis of [TetR+B0015]mini&dig XP

    • Digestion:[PompC+B0034]E first

    • Electrophoresis of [PompC+B0034]E

    • Digestion: [PompC+B0034]E, S later

    • Ligation: insert[PompC]ES+[B0034]XP/vector[PSB1K3]EP

    28

    • Electrophoresis of [PompC+B0034]ES not OK

    • Transformation of PompC+B0034+PSB1K3.

    • Electrophoresis of B0034+ho1

    • Digestion: [B0034+ho1]XP &[Pcons+RBS+pcyA]E

    29

    • PCR and electrophoresis test. Electrophoresis of digested B0034+ho1 &Pcons+RBS+pcyA.

    30

    • Cultivation of PompC+B0034+PSB1K3 E.coli in liquid LB-K tubes

    • Ligation: insert[TetR]ES+[B0015]XP/vector[PSB1K3]EP

    • Transformation of Plac+PSB1A3 on LB-A plate

    • Digestion:B0015 Ar dig XP, pSB1A3 Ar dig EP and pSB1C3 Cr dig EP

    • ligation:37。C RBS+mGFP+ter. [B0015], Cr

    • transformation of 37。C RBS+mGFP+ter., Cr

    31

    • Mini-prep of cultivated PompC+B0034 Kr E.coli, then

    • Digestion and electrophoresis of [PompC+B0034]E

    • Decide to determine the sequence of PompC+B0015.

    • Cultivation of ho1 with liquid LB-K and pSB1C3 with liquid LB-C.

    • PCR of 37。C RBS+mGFP+ter. Cr

    • electrophoresis of 37。C RBS+mGFP+ter., ter.[B0015] digXP, pSB1C3 dig EP and pSB1A3 dig EP

    • transformation of BBa _E1010 Kr

    June 2013

    Sunday

    Monday

    Tuesday

    Wednesday

    Thursday

    Friday

    Saturday

    1

    • PCR and single colony isolation of TetR+B0015+PSB1K3

    • Electrophoresis of PCR product made at this morning and Plac(5/30 Ar) OK

    • Digestion:[Ter(B0015)]XP

    • Transformation of Ter(B0015) on LB-A plate & mGFP on LB-K plate & 37oC RBS+mGFP+Ter(B0015) on LB-C plate

    • Ligation: insert 37oC RBS[ES] & Ter(B0015)[XP]/ vector pSB1C3[EP]

    • Single colony isolation from Ter(B0015) LB-A plate, and cultivation in liquid LB-A

    2

    • Ligation: insert [TetR]ES+[B0015]XP/vector [PSB1K3]EP and transformation of this part.

    • Ligation: B0032+ho1&B0034+ho1

    • Transformation of B0032+ho1&B0034+ho1

    • Cultivation of pSB1C3(C20&C10)with liquid LB.

    • PCR of insert fragment[37oC RBS+mGFP+Ter(B0015)]

    • Transformation of mRFP on LB-A plate& LB-C plate& LB-K plate-Mini-prep of cultivated Ter(B0015) E.coli.

    3

    • PCR of Plac &TetR+B0015

    • Mini-prep of pSB1C3

    • Cultivation of B0032+ho1&B0034+ho1 with LB-A plate PCR of B0032+ho1&B0034+ho1

    • Electrophoresis of B0032+ho1&B0034+ho1──NOT OK.

    • Single colony isolation from 37oC RBS+mGFP+Ter(B0015) LB-K plate, and cultivation in liquid LB-K & mRFP LB-A, and cultivation in liquid LB-K

    • Mini-prep of cultivated 37oC RBS+mGFP+Ter(B0015) E.coli

    • Digestion:[37oC RBS+mGFP+Ter(B0015)]XP

    4

    • Electrophoresis of Plac .TetR+B0015 OK

    • Cultivation of Plac.TetR+B0015 in liquid LB-A tubes. PCR of B0032+ho1&B0034+ho1

    • Electrophoresis of B0032+ho1(OK)&B0034+ho1(After PCR)

    • Mini-prep of B0032+ho1. Digestion: [B0032+ho1]XP.

    • Cultivation of ho1 with liquid LB-K

    • Single colony isolation from mRFP LB-A plate, and cultivation in liquid LB-A

    • Digestion:[37oC RBS+mGFP+Ter(B0015)]XP

    • PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)].

    5

    • Mini-prep of cultivated Plac and TetR+B0015 E.coli and then conduct digestion :[Plac]P&[TetR+B0015]P

    • Electrophoresis of [Plac ]P and [TetR+B0015]P,[Plac]XP and [TetR+B0015]XP

    • Ligation: insert[PompC+B0034]ES+[LacI+J61048]XP/vector [PSB1A3]EP

    • Electrophoresis of B0032+ho1 mini&B0032+ho1(digested)&pSB1C3 mini&pSB1C3(digested)&ho1 mini&ho1(digested)

    • Mini-prep of ho1

    • Digestion: [ho1]XP

    • Digestion:[37oC RBS+mGFP+Ter(B0015)]XP

    • Transformation of mRFP on LB-A plate

    6

    • Transformation of PompC+B0034+LacI+J61048+PSB1A3

    • Ligation: insert [PompC+B0034]ES+[LacI+J61048]XP/vector [PSB1A3]EP

    • Electrophoresis of B0032+ho1 mini&B0032+ho1(digested)&ho1 mini&ho1 (digested).

    • Single colony isolation from mRFP LB-A plate, and cultivation in liquid LB-A & LB-K & LB

    • Digestion:[Ter(B0015)]EX

    • Ligation: insert 37oC RBS+mGFP[ES]&Ter(B0015)[EX]

    • Transformation of 37oCRBS+mGFP+Ter(B0015) on LB-K plate & LB-A plate, mRFP on LB-A plate

    7

    • Transformation of PompC+B0034+LacI+J61048 again

    • Single colony isolation of TetR+B0015 Kr E.coli

    • Digestion:[LacI+J61048]ES. PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)]

    • Single colony isolation from 37oCRBS+mGFP+Ter(B0015) LB-A plate, and cultivation in liquid LB-A.

    8

    • PCR + single colony isolation of PompC+B0034+LacI+J61048 E.coli

    • Electrophoresis of PompC+B0034+LacI+J61048 OK

    • Ligation: insert [LacI+J61048]ES+[Plac]XP/vector[PSB1C3]EP

    • Transformation of LacI+J61048+Plac on LB-C plate

    • Cultivation of PompC+B0034+LacI+J61048 in LB-A tubes & TetR+B0015 in LB-K tube & PSB1A3 in LB-A tube &PSB1C3 in LB-C tube.

    • Mini-prep of cultivated 37oCRBS+mGFP+Ter(B0015) E.coli

    • Digestion:[ 37oCRBS+mGFP+Ter(B0015)]XP

    • PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)].

    9

    • Mini-prep from cultivated PompC+B0034+LacI+J61048 Eco.li&PSB1C3 E.coli

    • digestion : [PompC+B0034+LacI+J61048]ES&[PSB1C3]EP

    • Electrophoresis of [PompC+B0034+LacI+J61048] ES& [LacI+J61048]ES&[PSB1C3]EP.

    • Single colony isolation from 37oCRBS + mGFP+Ter(B0015)

    10

    • Mini-prep of cultivated 37oCRBS+mGFP+Ter(B0015) E.coli

    • Digestion:[ 37oCRBS+mGFP+Ter(B0015)]XP

    • Single colony isolation from 37oCRBS+mGFP+Ter(B0015) LB-A plate, and cultivation in liquid LB-A.

    11

    • PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)]

    • Transformation of mRFP on LB-K plate.

    12

    • Single colony isolation from mRFP LB-K plate, and cultivation in liquid LB-K

    13

    • Ligation: insert[PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1C3]EP and insert[TetR]ES+[B0015]XP/vector[PSB1C3]EP.

    • Mini-prep of cultivated mRFP E.coli

    • Digestion:[mRFP]ES & [Ter(J61048)]XP & [37OCRBS+mGFP]XP

    • Ligation: insert 37oCRBS+LuxR[ES]&37oCRBS+mGFP[XP] /vector pSB1A3[EP]

    • Transformation of 37oCRBS +LuxR+37oCRBS+mGFP on LB-A plate & mRFP +Ter(J61048) on LB-C plate& Ter(J61048) on LB-A plate.

    14

    • Transformation of PompC+B0034+LacI+J61048+PSB1C3 and TetR+B0015+PSB1C3

    • Cultivation of B0032+ho1 with liquid LB-A. PCR of insert fragment[Ter(B0015)]

    • Single colony isolation from Ter(J61048) LB-A plate, and cultivation in liquid LB-A & 37OCRBS+mGFP on LB-K plate, and cultivation in liquid LB-K

    • Transformation of 37oCRBS +LuxR+37oCRBS+mGFP on LB-A plate & mRFP +Ter(J61048) on LB-C plate

    • Mini-prep of cultivated Ter (J61048)& 37oCRBS+mGFP E.coli

    15

    • Digestion:[B0032]XP

    • Transformation of PompC+B0034+LacI+J61048 +PSB1C3and TetR+B0015 +PSB1C3 again

    • Ligation: insert[PompC]ES+[B0032]XP/vector[PSB1K3]EP, then transformation of this part

    • Mini-prep of B0032+ho1. PCR of insert fragment[37oCRBS + LuxR + 37oCRBS+mGFP]&[mRFP+Ter(J61048)].

    9

    • Mini-prep from cultivated PompC+B0034+LacI+J61048 Eco.li&PSB1C3 E.coli

    • digestion : [PompC+B0034+LacI+J61048]ES&[PSB1C3]EP

    • Electrophoresis of [PompC+B0034+LacI+J61048] ES& [LacI+J61048]ES&[PSB1C3]EP.

    • Single colony isolation from 37oCRBS + mGFP+Ter(B0015)

    10

    • Mini-prep of cultivated 37oCRBS+mGFP+Ter(B0015) E.coli

    • Digestion:[ 37oCRBS+mGFP+Ter(B0015)]XP

    • Single colony isolation from 37oCRBS+mGFP+Ter(B0015) LB-A plate, and cultivation in liquid LB-A.

    11

    • PCR of insert fragment[37oCRBS+mGFP+Ter(B0015)]

    • Transformation of mRFP on LB-K plate.

    12

    • Single colony isolation from mRFP LB-K plate, and cultivation in liquid LB-K

    13

    • Ligation: insert[PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1C3]EP and insert[TetR]ES+[B0015]XP/vector[PSB1C3]EP.

    • Mini-prep of cultivated mRFP E.coli

    • Digestion:[mRFP]ES & [Ter(J61048)]XP & [37OCRBS+mGFP]XP

    • Ligation: insert 37oCRBS+LuxR[ES]&37oCRBS+mGFP[XP] /vector pSB1A3[EP]

    • Transformation of 37oCRBS +LuxR+37oCRBS+mGFP on LB-A plate & mRFP +Ter(J61048) on LB-C plate& Ter(J61048) on LB-A plate.

    14

    • Transformation of PompC+B0034+LacI+J61048+PSB1C3 and TetR+B0015+PSB1C3

    • Cultivation of B0032+ho1 with liquid LB-A. PCR of insert fragment[Ter(B0015)]

    • Single colony isolation from Ter(J61048) LB-A plate, and cultivation in liquid LB-A & 37OCRBS+mGFP on LB-K plate, and cultivation in liquid LB-K

    • Transformation of 37oCRBS +LuxR+37oCRBS+mGFP on LB-A plate & mRFP +Ter(J61048) on LB-C plate

    • Mini-prep of cultivated Ter (J61048)& 37oCRBS+mGFP E.coli

    15

    • Digestion:[B0032]XP

    • Transformation of PompC+B0034+LacI+J61048 +PSB1C3and TetR+B0015 +PSB1C3 again

    • Ligation: insert[PompC]ES+[B0032]XP/vector[PSB1K3]EP, then transformation of this part

    • Mini-prep of B0032+ho1. PCR of insert fragment[37oCRBS + LuxR + 37oCRBS+mGFP]&[mRFP+Ter(J61048)].

    16

    • PCR + single colony isolation of PompC+B003+LacI+J61048 Cr E.coli&TetR+B0015 Cr E.coli&PompC+B0032 Kr E.coli, then Electrophoresis of these parts

    17

    • Mini-prep of cultivated PompC+B0034 E.coli and TetR+B0015 Cr E.coli

    18

    19

    20

    • Ligation: Pcons+RBS+pcyA&B0032+ho1&pSB1A3.

    21

    • Ligation: insert[PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1A3]EP

    • Transformation of Pcons+RBS+pcyA+B0032+ho1 on LB-A plate. PCR of insert fragment[37oCRBS + LuxR + 37oCRBS+mGFP]

    • Digestion:[pSB1C3]EP&[mRFP]ES&[Ter(J61048)]XP

    • Ligation: insert mRFP[ES]&Ter(J61048)[XP] /vector pSB1C3[EP]

    • Transformation of mRFP + Ter (J61048) on LB-C plate

    • Single colony isolation from 37oCRBS + LuxR +37oCRBS+mGFP LB-A plate, and cultivation in liquid LB-A.

    22

    • Transformation of PompC+B0034+LacI+J61048+PSB1A3 but there is no colony appearing. PCR of insert fragment[mRFP+Ter(J61048)]

    • Mini-prep of cultivated 37o CRBS + LuxR + 37oCRBS+mGFP E.coli

    • Digestion:[ 37o CRBS + LuxR + 37oCRBS+mGFP]ES

    23

    • Digestion:[PompC+B0032]ES and [TetR+B0015]XP,[LacI+J61048]XP

    • Ligation: insert [PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1C3]EP

    • Transformation of PompC+B0034+LacI+J61048 +PSB1C3

    • Electrophoresis of [PompC+B0032]ES & [TetR+B0015]XP

    • Digestion again:[PompC+B0032]ES&[LacI+J61048]XP&[LacI+B0015]XP

    • Transformation of mRFP + Ter (J61048) on LB-C plate

    24

    • PCR and single colony isolation of PompC+B0034+LacI+J61048+PSB1C3 E.coli

    • Electrophoresis of [B0032]dig ES&[TetR+B0015]dig XP&[LacI+J61048]dig XP&PCR product [PompC+B0034+LacI+J61048] ,[TetR+B0015] OK

    • Decide to determine the sequence of [TetR+B0015] OK.

    • Single colony isolation from 6/21 plate.

    • Cultivation of Pcons+ RBS+pcyA+B0032+ho1 with liquid LB-A.

    • Electrophoresis of Pcons+RBS+PcyA+B0032+ho1.

    • PCR of insert fragment[mRFP+Ter(J61048)].

    25

    • Ligation:insert[PompC+B0034]ES+[LacI+J61048]XP&[PompC]ES+[B0032]XP/vector[PSB1C3]EP and [PompC+B0034]ES+[LacI+J61048]XP/vector[PSB1A3]EP

    • Transformation of the ligation product made today

    • PCR of Pcons+RBS+pcyA+B0032+ho1.

    • Cultivation of Pcons+RBS+pcyA+B0032+ho1 with liquid LB-A

    • Electrophoresis of Pcons+RBS+pcyA+B0032+ho1.

    • Mini-prep of Pcons+RBS+pcyA+B0032+ho1.

    • Ligation: insert mRFP[ES]&Ter(J61048)[XP]/vector pSB1C3[EP]

    26

    • PCR and single colony isolation of PompC+B0034+LacI+J61048+PSB1A3& PompC+B0034+LacI+J61048+PSB1C3&PompC+B0032+PSB1C3 E.coli

    • Electrophoresis of the PCR product made today

    • Electrophoresis of Pcons+RBS+pcyA+B0032+ho1──NOT OK

    • Ligation: Pcons+RBS+pcyA&B0032+ho1

    • Transformation of Pcons+RBS+pcyA+B0032+ho1. Transformation of mRFP + Ter (J61048) on LB-C plate

    27

    • Cultivation of PompC+B0034+LacI+J61048+PSB1A3 E.coli in liquid LB-A tubes

    • PCR of Pcons+RBS+pcyA+B0032+ho1

    • Cultivation of Pcons+RBS+pcyA+B0032+ho1 with liquid LB-A

    • Electrophoresis of Pcons+RBS+pcyA+B0032+ho1──NOT OK

    • Transformation of mRFP + Ter (J61048) on LB-C plate

    28

    • PCR of Pcons+RBS+pcyA+B0032+ho1

    • Electrophoresis of Pcons+RBS+pcAa+B0032+ho1──NOT OK

    • Ligation: insert mRFP[ES]&Ter(J61048)[XP] /vector pSB1C3[EP]

    29

    • Cultivation of PompC+B0034+LacI+J61048+PSB1A3 E.coli in liquid LB-A tubes again.

    • Ligation: Pcons+RBS+pcyA&B0032+ho1&pSB1A3

    • Transformation of ligated Pcons+RBS+pcyA+B0032+ho1

    • Single colony isolation from 6/21 Pcons+RBS+pcyA+B0032+ho1 LB plate

    • PCR of 6/21 Pcons+RBS+pcyA+B0032+ho1

    • Electrophoresis of 621 Pcons+RBS+pcyA+B0032+ho1─NOT OK

    • Transformation of mRFP+Ter(J61048) on LB-C plate

    • Single colony isolation from 37oCRBS +LuxR +37oCRBS+mGFP LB-A plate, and cultivation in liquid LB-A

    30

    • Mini-prep of cultivated PompC+B0034+LacI+J61048+PSB1A3 E.coli and then digest with EcoR1 and SpeI:[ PompC+B0034+LacI+J61048]dig ES

    • Electrophoresis of [PompC+B0034+LacI+J61048+PSB1A3] mini and [ PompC+B0034+LacI+J61048]dig ES OK

    • Decide to determine sequence of[ PompC+B0034+LacI+J61048]

    • Cultivation of 6/29 Pcons+RBS+PcyA+B0032+ho1 LB plate with liquid LB-A

    • PCR of 6/29 Pcons+RBS+PcyA+B0032+ho1

    • Cultivation of 6/29 re-ligated Pcons+RBS+PcyA+B0032+ho1

    • PCR of 6/29 re-ligated Pcons+RBS+pcyA+B0032+ho1

    • Electrophoresis of all above─NOT OK

    July 2013

    Sunday

    Monday

    Tuesday

    Wednesday

    Thursday

    Friday

    Saturday

    1

    • Ligation: insert [B0030]ES+[ TetR+B0015]XP/vector [PSB1C3]EP

    • transformation of this part and cultivation of this part on LB-C plate

    • Digestion : mRFP [EP]

    • Ligation : insert mRFP [EP]/vector pSB1C3 [EP]

    • Transformation of mRFP ,and cultivation on LB-C plate

    2

    • PCR +single colony isolation B0030+TetR+B0015 Cr E.coli

    • Single colony isolation from mRFP LB-C plate, and cultivation of in liquid LB-C

    3

    • Electrophoresis of B0030+TetR+B0015 not OK

    • electrophoresis of B0030+TetR+B0015 again OK

    • Ligation: Pcons+B0030+pcyA+B0032+ho1

    • Transformation of Pcons+B0030+pcyA+B0032+ho1

    • Mini-prep of cultivated 37℃RBS+luxR+37℃RBS+mGFP and mRFP E.coli

    • Digestion : mRFP [ES] & pSB1A3 [EP] & pSB1C3 [EP] & pSB1K3 [EP]

    • Electrophoresis of insert fragment [mRFP & pSB1A3 & pSB1C3 & pSB1K3]---- OK

    4

    • Cultivation of B0030+TetR+B0015+PSB1C3 in liquid LB-C tubes. PCR of Pcons+B0030+pcyA+B0032+ho1

    • Electrophoresis of Pcons+B0030+pcyA+B0032+ho1

    • Cultivation of Pcons+B0030+pcyA+B0032+ho1 with liquid LB-A

    • Ligation : insert mRFP [ES] & J61048 [XP]/ vector pSB1K3 [EP]

    • Transformation of mRFP+J61048 ,and cultivation on LB-K plate

    5

    • Mini-prep of cultivated B0030+TetR+B0015+PSB1C3 Cr E.coli and then digest with XbaI and Pst1

    • Electrophoresis of[ B0030+TetR+B0015]mini&dig XP

    • Decide to determine the sequence of B0030+TetR+B0015

    • Digestion: [Pcons+B0030+pcyA+B0032+ho1]ES

    • Electrophoresis of Pcons+B0030+pcyA+B0032+ho1──NOT OK

    • PCR of insert fragment [mRFP+J61048]

    6

    • Ligation :insert [PompC]ES+[B0032]XP/vector [PSB1C3] EP and insert [LacI+J61048]ES&[Plac]XP/vector [PSB1C3]EP

    • Transformation of PompC+B0030+PSB1C3 and LacI+J61048+Plac+PSB1C3

    • Ligation : insert 37℃RBS+luxR+37℃RBS+mGFP [ES] & B0015 [XP]/ vector pSB1C3 [EP]

    • Transformation of 37℃RBS+luxR+37℃RBS+mGFP+B0015 ,and cultivation on LB-C plate

    • Electrophoresis of insert fragment [mRFP+J61048]----undetermined

    7

    8

    • PCR + single colony isolation of LacI+J61048+Plac +PSB1C3 E.coli

    • Electrophoresis of LacI+J61048+Plac OK

    9

    • Single colony isolation from mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K

    • Mini-prep of cultivated mRFP+J61048 E.coli

    • Digestion : mRFP+J61048 [XP]

    10

    • Cultivation of LacI+J61048+Plac in liquid LB-C tubes.

    • Electrophoresis of insert fragment [mRFP+J61048]----NOT OK

    11

    • Mini-prep of cultivated LacI+J61048+Plac E.coli

    12

    • Ligation again:Insert [PompC]ES&[B0032]XP/vector [PSB1K3]EP

    • Digestion: [LacI+J61048+Plac]XP

    • Electrophoresis of [LacI+J61048+Plac]mini&dig XP OK

    • Decide to determine the sequence of LacI+J61048+Plac

    • Transformation of PompC(Ar)+B0032(Ar)+PSB1K3 again but there is still no colony

    • Transformation of 37℃RBS+luxR+37℃RBS+mGFP+B0015, and cultivation on LB-C plate

    • Transformation of mRFP+J61048 ,and cultivation on LB-K plate.

    13

    • Ligation: Insert [PompC]ES&[B0032]XP/vector [PSB1K3]EP. PCR of insert fragment [37℃RBS+luxR+37℃RBS+mGFP+B0015 & mRFP+J61048].

    14

    • Transformation of the ligation product made yesterday on LB-K plate but there is no colony appearing

    • Electrophoresis of insert fragment [37℃RBS+luxR+37℃RBS+mGFP+B0015]----OK

    • Electrophoresis of insert fragment [mRFP+J61048]----NOT OK RBS+mGFP+B0015 LB-C plate, and cultivation of in liquid LB-C. PCR of insert fragment [mRFP+J61048]----NOT OK

    15

    • Ligation and trans formation of insert [ B0030]ES+ [TetR+B0015]XP/vector [PSB1C3]EP and cultivation on LB-C plate

    • Transformation of PSB1K3 for testing

    • Transformation of Pcons

    • Digestion: [B0030+pcyA]ES

    • Electrophoresis of Pcons──OK

    • Electrophoresis of insert fragment [mRFP+J61048]----NOT OK

    • Mini-prep of cultivated 37℃RBS+luxR+37℃RBS+mGFP+B0015 E.coli----NOT OK

    • PCR of insert fragment [mRFP+J61048]----OK

    16

    • Digestion:[PompC]ES,[PSB1K3]&[PSB1C3]EPand Electrophoresis for checking these parts OK

    • PCR + singles colony isolation of B0030+TetR+B0015 +PSB1C3 E.coli and electrophoresis of this insert fragment not OK Because the PCR electrophoresis result is strange，we conduct this experience again OK

    • Ligation: Pcons&Pcons+B0030+pcyA+B0032+ho1&pSB1K3.

    • Transformation of Pcons+B0032+pcyA+B0030+ho1.

    • Cultivation of Pcons with liquid LB-C

    • Electrophoresis of Pcons──NOT OK

    • Single colony isolation from 37℃RBS+luxR+37℃RBS+mGFP+B0015 LB-C plate, and cultivation of in liquid LB-C

    • Single colony isolation from mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K

    17

    • Transformation of PompC+B0032+PSB1C3 and Cultivation of B0030+TetR+B0015 Cr E.coli in liquid LB-C tube.

    • Transformation of Pcons

    • transformation of tetR plasmid and cultivation on LB –A plate 3. Digestion: ter. [J61048], ter. [B0015] dig EP

    • Digestion : pSB1C3 [ES] & pSB1C3 [XP]

    • Ligation : insert sRNA+target 1 [ES]/vector pSB1C3 [ES]

    • Transformation of 37℃RBS+luxR+37℃RBS+mGFP+B0015 & sRNA 1 & sRNA 2 & target 1 & target 2 ,and cultivation on LB-C plate

    • Transformation of mRFP+J61048 & sRNA+target 1 & sRNA+target 2 ,and cultivation on LB-K plate

    • Electrophoresis of insert fragment [37℃RBS+luxR+37℃RBS+mGFP+B0015 & mRFP+J61048 & pSB1A3 & pSB1C3 & pSB1K3]-- mRFP+J61048 NOT OK Digestion : mRFP+J61048 [XP]

    18

    • Mini-prep of cultivated B0030+TetR+B0015 Cr E.coli and then digest these mini with XbaI and PstI

    • PCR + single colony isolation of PompC+B0032 Cr E.coli and electrophoresis of this insert fragment OK Cultivation of [PompC+B0032] Cr E.coli in liquid LB-C Tubes.

    • Cultivation of Pcons with liquid LB-C

    • Digestion : pSB1C3 [ES] & pSB1C3 [XP]

    • Mini-prep of Pcons. Electrophoresis of Pcons──NOT OK

    • Four colonies isolation from Pcons LB plate and use streak plate method to isolate the pure cultures. Digestion : mRFP+J61048 [XP]

    • Single colony isolation from sRNA+target 1 & sRNA+target 2 LB-K plate, and cultivation of in liquid LB-K

    • Single colony isolation from Plux LB-A plate, and cultivation of in liquid LB-A----NOT OK

    19

    • Mini-prep of cultivated PompC+B0032 Cr E.coli, And then digest with EcoR1 and SpeI

    • Electrophoresis of PSB1K3 made at 7/16 and B0030+TetR+B0015, PompC+B0032 OK.

    • Cultivation of Pcons with liquid LB-C

    • Single colony isolation from37℃RBS+luxR+37℃RBS+mGFP+B0015 LB-C plate, and cultivation of in liquid LB-C

    • Single colony isolation from mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K(?)

    • PCR of insert fragment [sRNA 1 & sRNA 2 & target 1 & target 2]

    • Electrophoresis of insert fragment [sRNA 1 & sRNA 2 & target 1 & target 2]---- target 1?

    • Mini-prep of cultivated sRNA+target 1 & sRNA+target 2 E.coli.

    20

    • Mini-prep of Pcons. -Digestion: [Pcons]ES.

    • Electrophoresis of Pcons──NOT OK.

    • Single colony isolation from sRNA 1 & sRNA 2 & target 2 LB-C plate, and cultivation of in liquid LB-C

    • Single colony isolation from mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K

    • Transformation of Plux ,and cultivation on LB-A plate

    • Single colony isolation from Plux & PLac & Pcons LB-A plate, and cultivation of in liquid LB-A

    • Transformation of RBS+lacI+B0010+B0012+pLac ,and cultivation on LB-K plate

    • Mini-prep of cultivated 37℃RBS+luxR+37℃RBS+mGFP+B0015 & mRFP+J61048 & sRNA 1 & sRNA 2 & target 2 E.coli

    21

    • Single colony isolation from 7/15 Pcons LB plate(Red one).

    • Cultivation of Pcons with LB-C. PCR of insert fragment [target 1]

    • Transformation of RBS+lacI+B0010+B0012+pLac ,and cultivation on LB-K plate

    • Electrophoresis of insert fragment [37℃RBS+luxR+37℃RBS+mGFP+B0015 & mRFP+J61048 & sRNA 1 & sRNA 2 & target 1 & target 2]----?

    • Mini-prep of cultivated Plux & PLac E.coli Digestion : Plux [ES] & Plac [ES] & mRFP+J61048 [XP].

    22

    • Ligation :insert [LacI+J61048]ES&[Plac]XP/vector [PSB1C3] EP

    • Transformation of the ligation product made at this morning and cultivation on LB-C plate.

    • Mini-prep of Pcons+mRFP

    • Digestion: [Pcons]ES.

    • Electrophoresis of Pcons+mRFP──NOT OK.

    • Cultivation of Pcons+mRFP with liquid LB-C. PCR of Pcons+mRFP

    • Electrophoresis of Pcons+mRFP──OK.

    • Electrophoresis of insert fragment [Plux & Plac & mRFP+J61048?]----?

    23

    • PCR +Single colony isolation of LacI+J61048+Plac and electrophoresis Of this insert fragment OK

    • Mini-prep of Pcons-mRFP.

    • Digestion: [B0030]XP&[B0032]XP&[B0023]XP&[Pcons]ES

    • Electrophoresis of B0030,B0032,B0034,Pcons──OK

    • Transformation of B0030+pcyA+B0032+ho1+B0030/B0032/B0034

    • Electrophoresis of insert fragment [mRFP+J61048]----?

    • Transformation of RBS+lacI+B0010+B0012+Plac ,and cultivation on LB-K plate----NOT OK

    • Single colony isolation from RBS+lacI+B0010+B0012+pLac & mRFP+J61048 LB-K plate, and cultivation of in liquid LB-K

    24

    • Cultivation of LacI+J61048+Plac E.coli in liquid LB-C tubes

    • Mini-prep of cultivated LacI+J61048+Plac E.coli.

    • Cultivation of B0030+pcyA+B0032+ho1+B0032 with liquid LB

    • Transformation of B0030+pcyA+B0032+ho1+B0030/B0032/B0034

    • Mini-prep of cultivated mRFP+J61048 E.coli

    • Digestion : mRFP+J61048 [XP]

    • Transformation of target 1 ,and cultivation on LB-C plate

    • Single colony isolation from RBS+lacI+B0010+B0012+pLac plate, and cultivation of in liquid LB-without antibiotics----NO E.coli?

    25

    • Digestion of LacI+J61048+Plac Cr [XP]

    • Electrophoresis of[ LacI+J61048 +Plac]dig XP OK

    • Decide to determine the sequence of LacI+J61048 +Plac

    • Ligation :insert [B0030]ES& [TetR+B0015]XP/vector [PSB1K3]EP.

    • Cultivation of B0030+pcyA+B0032+ho1+B0030/B0032/B0034 with liquid LB.

    • PCR of insert fragment [target 1]

    • Single colony isolation from RBS+lacI+B0010+B0012+pLac plate , and cultivation of in liquid LB-without antibiotics----NO E.coli?

    • Electrophoresis of insert fragment [mRFP+J61048 & target 1]----?

    26

    • Transformation of B0030+TetR+B0015+PSB1K3 and cultivation on LB-K plate. -Mini-prep of B0030+pcyAB0032+ho1+B0030/B0032/B0034.

    • Digestion: [B0030+pcyA+B0032+ho1+B0030/B0032/B0034]XP

    • Electrophoresis of digested B0030+pcyA+B0032+ho1+B0030/B0032/B0034──OK

    • Electrophoresis of B0030,B0032,B0034,Pcons──OK

    • Mini-prep of cultivated target 1 & mRFP+J61048 E.coli

    • Digestion: target 1 [ES] & mRFP [XP]?

    • Electrophoresis of insert fragment [RBS+lacI+B0010+B0012+pLac & mRFP & target 1]----?

    27

    • PCR and single colony isolation of B0030+TetR+B0015 Kr E.coli

    • Cultivation of B0030+TetR+B0015 in liquid LB-tube.

    • Ligation: pSB1C3&Pcons&B0030+pcyA+B0032+ho1+B0030

    28

    • Cultivation of B0030+TetR+B0015 in liquid LB-K tubes again

    • Ligation:insert[PompC+B0032]ES+[LacI+J61048+Plac]XP/vector[PSB1K3]EP

    • Mini-prep of cultivated B0030+TetR+B0015 Kr E.coli

    • Transformation of Pcons+B0030+pcyA+B0032+ho1+B0030

    29

    • Transformation of PompC+B0032+LacI+J61048+Plac+PSB1K3. -Transformation of Cph8.

    • Digestion: [Pcons]XP.

    30

    • Ligation:insert[PompC+B0032]ES+[LacI+J61048+Plac]XP/vector[PSB1K3]EP&[PSB1A3]EP

    • Transformation of PompC+B0032+LacI+J61048+Plac+PSB1K3&PSB1A3.

    • Electrophoresis of Pcons ─OK. Digestion: [Pcons]SP.

    31

    • PCR + single colony isolation of PompC+B0032+LacI+J61048+Plac+PSB1K3&PSB1A3 E.coli

    • Electrophoresis of PompC+B0032+LacI+J61048+Plac OK.

    • Ligation: Pcons&B0030+pcyA+B0032+ho1+B0030.

    • Transformation of Pcons+B0030+pcyA+B0032+ho1+B0030

    August 2013

    Sunday

    Monday

    Tuesday

    Wednesday

    Thursday

    Friday

    Saturday

    1

    • Cultivation of PompC+B0032+LacI+J61048+Plac Kr&Ar E.coli in liquid LB-K&LB-A tubes.

    • Cultivation of Pcons+B0030+pcyA+ B0032+ho1+B0030 with liquid LB-K

    2

    • Mini-prep of cultivated PompC+B0032+LacI+J61048+Plac Kr&Ar E.coli

    • Digestion: [PompC+B0032+LacI+J61048]ES. Mini-prep of Pcons+B0030+pcyA+ B0032+ho1+B0030.

    • Digestion: [Pcons+B0030+pcyA+ B0032+ho1+B0030]ES

    • Electrophoresis of Pcons+B0030+pcyA+ B0032+ho1+B0030──OK

    3

    4

    5

    6

    7

    8

    9

    • Ligation: insert[PompC+B0032]ES+[LacI+J61048+Plac]XP/vector[PSB1K3]EP, then transformation of this ligation product

    10

    11

    • PCR + single colony isolation of PompC+B0032+LacI+J61048+Plac Kr E.coli

    • Electrophoresis of PompC+B0032+LacI+J61048+Plac not OK

    12

    • Colony PCR of 37。C RBS+LuxR+37。C RBS+mGFP Ar E.coli&Plux+sRNA2 Kr E.coli&mRFP+J61048 Kr E.coli

    • Electrophoresis of [target site 1]PCR&[37。C RBS+LuxR+37。C RBS+mGFP]PCR&[LacI+J61048+Plac+sRNA2]dig XP&[target site2+mRFP]dig ES&[Plux+sRNA2]PCR&[mRFP+J61048]PCR

    • Cutivation of Plux+sRNA2 Kr E.coli in liquid LB-K tube

    13

    • Mini-prep of cultivated Plux+sRNA2 Kr E.coli(fault)

    • Transformation of 37。C RBS+LuxR+37。C RBS+mGFP in PSB1A3

    • Ligation:insert[sRNA+target site1]XP/vector[PSB1C3]XP Colony PCR of target site2+mRFP Ar E.coli

    • Cultivation of Plux+sRNA2 Kr E.coli in liquid LB-K tube

    14

    • Mini-prep of cultivated Plux+sRNA2 Kr E.coli

    • Digestion:[Plux+sRNA2]XP

    • Digestion:[Plux+sRNA2]XP

    • Colony PCR of 37。C RBS+LuxR+37。C RBS+mGFP Ar E.coli

    • Electrophoresis of [target site2+mRFP]PCR

    • Transformation of target site1 in PSB1C3

    • Electrophoresis of [target site2+mRFP]PCR&[LacI+J61048+Plac+sRNA2]mini

    15

    • Electrophoresis of [LacI+J61048+ Plac+sRNA2]dig XP&[Plux+sRNA2]dig XP&[37。C RBS+LuxR+37。C RBS+mGFP]PCR

    • Colony PCR of target 1 Cr E.coli&mRFP+J61048 Kr E.coli and then electrophoresis of these PCR products

    16

    • Colony PCR of target 1 Cr E.coli and then electrophoresis of PCR products

    • Transformation of mRFP+J610148 in PSB1K3(failed)

    17

    • Colony PCR of mRFP+J61048 Kr E.coli and then electrophoresis of the PCR product

    • Colony PCR of target 1 Cr E.coli and then electrophoresis of the PCR product

    • Transformation of mRFP+J610148 in PSB1K3(failed)

    • Cultivation of mRFP+J61048 Kr E.coli in liquid LB-K tube(failed)

    18

    • Colony PCR of mRFP+J61048 Kr E.coli

    • Digestion:[J61048]XP&[target site1]XP

    • Ligation:insert[target site2]ES+[J61048]XP/vector[PSB1C3]EP

    • Electrophoresis of [mRFP+J61048]PCR&[target site1]PCR

    19

    • Cultivation of mRFP+J61048 Kr E.coli in liquid LB-K tube

    • Transformation of target site2+mRFP+J61048 in PSB1C3

    • Ligation:insert[target site 1]/vector[PSB1C3]

    • Mini-prep of cultivated mRFP+J61048 Kr E.coli

    • Digestion:[mRFP+J61048]XP

    20

    • Colony PCR of target site2+mRFP+J61048 Cr E.coli

    • Transformation of target site1 in PSBC3 Cr E.coli

    • Electrophoresis of[mRFP+J61048]dig XP&[Plux+sRNA]digXP

    • Mini-prep of cultivated Plux+sRNA2 Kr E.coli Digestion:[Plux+sRNA2]XP

    21

    • Electeophoresis of [mRFP+J61048]dig XP&[ target site2+mRFP+J61048]PCR

    22

    • Cultivation of target site2+mRFP+J61048 Cr E.coli

    • Transformation of target site1 in PSB1C3

    • Ligation:insert[mRFP]ES+[J61048]XP/vector[PSB1K3]EP

    • Dig:LuxI(BBa_C0061)Transformation of mRFP+J61048 in PSb1K3&LuxI inPSB1A3

    • Mini-prep of cultivated target site2+mRFP+J61048 Cr E.coli and then digestion:[target site2+mRFP+J61048]XP

    23

    • Electrophoresis of [target site 2+mRFP+J61048]dig XP&[37。C RBS+LuxR+37。C RBS+GFP]PCR&[Pompc+B0032+LacI+J61048+Plac]PCR&[mRFP+J61048]PCR&[target site1]PCR

    • Cultivation of LuxI Ar E.coli in liquid LB-A tube

    • Colony PCR of target site1 Cr E.coli&mRFP+J61048Kr E.coli

    • Dig:B0034+LuxI(BBa_C0261)Transformation of B0034+LuxI in PSB1A3

    24

    • Cultivation of 37。C RBS+LuxR+37。C RBS+mGFP Ar E.coli&B0034+LuxI Ar E.coli in liquid LB-A tubes and Pompc+B0032+LacI+J61048+Plac Kr E.coli in liquid LB-K tube

    • Digestion:[37。C RBS+mGFP]ES&[37。C RBS+LuxR]XP&[sRNA1]ES&[Pcons]XP

    • ligation:insert[target site 1]XP/vector[PSB1C3]XP&insert[mRFP]ES+[B0015]XP/vector[PSB1C3]EP&insert[Pcons]ES+[target site 2+mRFP+J61048]XP/vector[PSB1C3]EP&insert[37。C RBS+mGFP]ES+[37。C RBS+LuxR]XP/vector[PSB1C3]EP&insert[sRNA1]ES+[Pcons]XP/vector[PSB1K3]EP&insert[Pcons]ES+[32。C RBS+mGFP]XP/vector[PSB1C3]EP and then transformation

    • mini-prep of cultivated LuxI Ar E.coli&B0034+LuxI Ar E.coli&Pompc+B0032+LacI+J61048+Plac Kr E.coli&37。C RBS+LuxR+37。C RBS+mGFP Ar E.coli

    25

    • Digestion:[LuxI]XP&[B0034+LuxI]ES&[ Pompc+B0032+LacI+J61048+Plac]ES&[7。C RBS+LuxR+37。C RBS+mGFP]&[PSB1A3.PSB1C3.PSB1K3]EP&[PSB1C3]ES and XP&[37。C RBS+mGFP]EP and then electrophoresis

    • Ligation:insert[B0034+LuxI]ES+[B0015]XP/vector[PSB1C3]EP&[Pcons]ES+[target site 2+mRFP+J61048]XP+[37。C RBS+mGFP]EP

    • Transformation of B0034+LuxI+B0015 inPSB1C3&Pcons+target site2+mRFP+J61048&Pcons+32。C RBS+mGFP in PSB1C3

    • Colony PCR of sRNA1+Pcons Kr E.coli&target site1 Cr E.coli&mRFP+B0015 Cr E.coli&37。C RBS+mGFP+37。C RBS+LuxR Cr E.coli

    • Electrophoresis of [target site1]PCR&[mRFP+B0015]PCR&[37。C RBS+mGFP+37。C RBS+LuxR]PCR+[sRNA1+Pcons]PCR

    26

    • Mini-prep of cultivate target site1Cr E.coli&mRFP+B0015 Cr E.coli&37。C RBS+mGFP+37。C RBS+LuxR Cr E.coli&sRNA+Pcons Kr E.coli

    • Digestion:[target site 1]ES&[ mRFP+B0015]XP&[37。C RBS+mGFP+37。C RBS+LuxR]ES&[ sRNA+Pcons]ES&[37。C　RBS+LuxR+37。C RBS+mGFP]ES

    • Colony PCR of B0034+LuxI+B0015 Cr E.coli&Pcons+target site 2+mRFP+J61048 Kr E.coli

    • Electrophoresis of those digestion and PCR products made today

    27

    • Mini-prep of cultivated Pcons+target site2+mRFP+J61048 K r E.coli

    • Digestion:[ Pcons+target site2+mRFP+J61048]ES&[Pompc+B0032+LacI+J61048+Plac]XP

    • Ligation insert[target site 1]ES+[mRFP+B0015]XP/vector[PSB1A3]EP&insert[Pcons+target site2+mRFP+J61048]ES+[Pcons+B0032+LacI+J61048+Plac+sRNA2]XP/vector[PSB1A3]EP&insert[Pcons]ES+[37。C RBS+mGFP+37。CRBS+LuxR]XP/vector[PSB1K3]EP&insert[Pcons]ES+[37。C RBS+LuxR+37。C RBS+mGFP]XP/vector[PSB1K3]EP&insert[Pcons+target site 2+mRFP+J61048]ES+[Plux+sRNA2]XP/vector[PSB1A3]EP&insert[Pompc+B0032+LacI+J61048+Plac]ES+[37。C RBS+mGFP+37。C RBS+LuxR]XP/vector[PSB1A3]EP&insert[Pompc+B0032+LacI+J61048+Plac]ES+[37。CＲＢＳ＋LuxR+37。C RBS+mGFP]XP/vector[PSB1C3]EP&insert[sRNA1(With Plux)]ES+[Pompc+B0032+LacI+J61048+Plac]XP/vector[PSB1A3]EP&insert[Pompc+B0032+LacI+J61048+Plac]ES+[target site2+mRFP]XP/vector[PSB1A3]&insert[Pcons+target site 2+mRFP+J61048]ES+[Pcons+B0032+LacI+J61048+Plac+sRNA2]XP/vector[mGFP]EP&insert[Pcons+target site2+mRFP+J61048]ES+[Plux+sRNA]XP/vector[mGFP]EP

    • Transformation of these ligation product and B0034+LuxI+B0015 in PSB1C3

    • Electrophoresis of [Pcons+tar get site 2+mRFP+J61048]PCR&[PSB1K3]dig EP&[Pcons+target site2+mRFP+J61048]dig ES&[Pompc+B0032+LacI+J61048+Plac]XP&[37CRBS+mGFP+37C

    28

    • Cultivation of B0034+LuxI+B0015 E.coli on LB-C plate&Pcons+37。C RBS+mGFP+37。C RBS+LuxR E.coli on LB-K plate&Pcons+37。C RBS+LuxR+37。C RBS+mGFP E.coli on LB-K plate&sRNA1(with Plux)+Pompc+B0032+LacI+J61048+Plac on LB-A plate

    • Colony PCR of target site 1+mRFP+B0015 Ar& Pcons+target site2+mRFP +J61048+Pcons+B0032+LacI+J61048+Plac+sRNA2 Ar & Pcons+37。C RBS+mGFP+37。C RBS+LuxR Kr&Pcons+target site 2+mRFP+J61048+Plac+sRNA2 Ar&Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR Ar&PomPC +B0032+LacI+J61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP Cr &sRNA2(with Plac)+Pompc+B0032+LacI+J61048+Plac Ar& Pompc+B0032+LacI+J61048+Plac+target site 2+mRFP+J61048 Ar E.coli and then electrophoresis of these PCR products

    29

    • Mini-prep of cultivated target site 1+mRFP+B0015 Ar &Pcons+target site 2+mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA 2 Ar&Pcons+37。C RBS＋mGFP+37。C RBS+LuxR Kr & Pompc+B0032+LacI+J61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP Cr & Pcons+target site 2+mRFP+J61048+Plux+sRNA2 Kr &Pcons+37。C RBS+LuxR+37。C RBS+mGFP Kr E.coli

    • Digestion:[ target site 1+mRFP+B0015]XP&[Pcons+target site 2+mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA 2]XP&[ Pcons+37。C＋mGFP+37。C RBS+LuxR ]ES&[ Pompc+B0032+LacI+J61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP]ES and then electrophoresis of these digestion products

    • Colony PCR of Pcons+target site mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA2 Ar &Pcons+target site2+mRFP+J61048+Plac+sRNA2 Ar &Pompc+B0032+LacI+J61048+Plac+37。 C RBS+mGFP+37。C RBS+LuxR Ar &Pompc+B0032+LacI+J61048+Plac+target site2+mRFP+J61048 Ar &sRNA1(with Plac)+Pompc+B0032+LacI+J61048+Plac Ar &Pcons+37。C RBS+LuxR+37。C RBS+mGFP Kr &B0034+LacI+B0015 Cr E.coli and then electrophoresis of these PCR products

    • Ligation:insert[Pcons]ES+[37。 C RBS+mGFP]XP/vector[PSB1C3]EP&insert[37。C RBS]ES+[LacI+J61048+Plac+sRNA2]XP/vector[PSB1K3]EP&insert[B0034+LuxI]ES+[J61048]

    30

    • Cultivation of target site1+mRFP+B0015 Ar and Pcons+target site2+mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA2 Ar E.coli in liquid LB-A tubes &Pcons+37。C RBS+mGFP+37。C RBS+LuxR Kr E.coli in liquid LB-K tube &Pompc+B0032+LacI+J+61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP Cr E.coli in liquid LB-C tube &Plux+sRNA 2 in liquid LB tube

    • Mini-prep of those cultivated E.coli which was cultivated this morning

    • Electrophoresis of the mini-prep products made today and [Pcons+target site 2+mRFP+J61048+Pcons+B0032+LacI+J61048+Plac+sRNA 2]dig ES &[ Pcons+target site 2+mRFP+J61048+Plux+sRNA2]dig ES&[cons+37。C RBS+LuxR+37。C RBS+mGFP]XP

    • digestion:[B0034]XP&[target site 2]XP colony PCR of Pcons+37。C RBS Cr E.coli&37。C RBS+LacI+J61048+Plac+sRNA2 Kr E.coli&B0034+LacI+J61048 Kr E.coli&B0032+LacI Kr E.coli and then electrophoresis of these PCR products

    31

    • Ligation:insert[Pcons]ES+[target site 2]XP/vector[PSB1K3]EP&insert[Pcons]ES+[B0034]XP/vector[PSB1K3]EP

    • Transformation of Pcons+target site 2 in PSB1K3&Pcons+B0034 in PSB1K3 & BBa_K516030(mRFP protein generator) in PSB1C3&BBa_K516132(Constitutive promoter [J23101] with mRFP, RBS B0032) in PSB1C3&Pcons+LacI+Plac+sRNA2 in PSB1K3

    • Mini-prep of cultivated Pcons+37。C RBS+mGFP Cr E.coli&37。C RBS+LacI+J61048+Plac+sRNA2 Cr E.coli&B0032+LuxI Kr E.coli

    • digestion:[ Pcons+37。C RBS+mGFP]ES&[37。C RBS+LacI+J61048+Plac+sRNA2]XP&[ B0032+LuxI]ES &[Pcons+37。C RBS+mGFP+37。C RBS+LuxR]ES&[Pompc+B0032+LacI+J61048+Plac+37。C RBS+LuxR+37。C RBS+mGFP]ES

    • cultivation of Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR Ar &Pompc+B0032+LacI+J61048+Plac+target site 2+mRFP=J61048 Ar E.coli on LB-A plates

    September 2013

    Sunday

    Monday

    Tuesday

    Wednesday

    Thursday

    Friday

    Saturday

    1

    • Ligation:insert[Pcons+37。C RBS+mGFP]ES+[37。C RBS+LacI+J61048+Plac+sRNA2]XP/vector[PSB1A3]EP&inert[Pcons+37。C RBS+mGFP]ES+[37。C RBS+LuxR]XP/vector[PSB1A3]EP

    2

    • Mini-prep of cultivated Pcons+37。C RBS+mGFP+37。C RBS+LacI+J61048+Plac+sRNA2 Ar E.coli Colony PCR of Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR Ar &sRNA(with Plux)+Pompc+B0032+LacI+J61048+Plac Ar &Pompc+B0032+LacI+J61048+Plac+target site2+mRFP+J61048 Ar &Pcons+37。C RBS+mGFP+37。C RBS+LuxR Ar E.coli Electrophoresis of PCR products today

    3

    • Mini-prep of cultivated Pcons+37。C RBS+mGFP+37。C RBS+LacI+J61048+Plac+sRNA2 Ar E.coli Colony PCR of Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR Ar &sRNA(with Plux)+Pompc+B0032+LacI+J61048+Plac Ar &Pompc+B0032+LacI+J61048+Plac+target site2+mRFP+J61048 Ar &Pcons+37。C RBS+mGFP+37。C RBS+LuxR Ar E.coli Electrophoresis of PCR products today

    • Digestion:[ Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP+37。C RBS+LuxR]ES&[ sRNA1(with Plux)+Pompc+B0032+LacI+J61048+Plac]ES&[ Pompc+B0032+LacI+J61048+Plac+target site2+mRFP+J61048]ES&[Pcons+37。C RBS+mGFP+37。C RBS+Lux]ES&[BBa_K516030(Constitutive promoter [J23101] with mRFP, RBS B0030) ]XP and then electrophoresis

    • Ligation:insert[Pcons]ES+[target site 2]XP/vector[PSB1K3]&insert[Pcons]ES+[B0034]XP/vector[PSB1K3]EP&insert[B0034+LuxI]ES+[B0015]XP/vector[PSB1C3]EP

    4

    • Colony PCR of Pcons+B0034 Kr&Pcons+target site Kr&Pcons+K516030 Kr&B0034+LacI+B0015 Cr E.coli and then electrophoresis of these PCR products

    • Ligation: insert[Pcons+B0032]ES+[LacI+J61048+Plac+sRNA2]XP/vector[PSB1K3]&insert[37。C RBS]ES+[LacI+J61048+Plac+sRNA2]XP/vector[PSB1K3]EP&insert[Pompc+B0032+LacI+J61048+Plac]ES+[37。C RBS+mGFP]XP/vector[PSB1C3]EPBackbone changing:[37。C RBS+mGFP]XP&[37。C RBS+LuxR]XP&[37。C RBS+LuxR+37。C RBS+mGFP]XP&[Pcons+B0030+PcyA+B0032+LacI+B0030]ES&[Pcons+target site2+mRFP+J61048]ES /vector[PSB1C3]ES and [sRNA+target site2]EP/vector[PSB1C3]EP

    • Digestion:[sRNA+target site2]EP Transformation of Pcons+B0032+LacI+J61048+Plac+sRNA2 in PSB1K3&37。C RBS+LacI+J61048+Plac+sRNA2 in PSB1K3&Pompc+B0032+LacI+J61048+Plac+37。C RBS+mGFP in PSB1C3&37。C RBS+mGFP in PSB1C3&37。C RBS+LuxR in PSB1C3&37。C RBS+LuxR+37。C RBS+mGFP in PSB1C3&Pcons+B0034+PcyA+B0032+LacI+B0030 in PSB1C3&Pcons+target site2+mRFP+J61048 in PSB1C3&sRNA+target site 2 in PSB1C3

    • Mini-prep of cultivated Pcons+B0034 Kr E.coli&Pcons+target site2 Kr E.coli&Pcons+K516030 Kr E.coli and then digestion:[ Pcons+B0034]ES and E&[ Pcons+target site2]ES and E&[Pcons+K516030]ES

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    Retrieved from "http://2013.igem.org/Team:NCTU_Formosa/notes"