Revision as of 06:55, 27 September 2013

Maestro E.coli

Notebook

RandomPromtoer-B0034-mRFP1-B0015

Start
PCR
- Tue Jun 11
B0034-mRFP1-B0015
100up, 200dn, KOD Plus Neo
Get K1084007 from our stock
- Tue Jun 11
PCR purification
- Wed Jun 12
B0034-mRFP1-B0015
Digestion
- Wed Jun 12
Cut K1084007 with SpeI, PstI (using 10×H buffer)
Cut B0034-mRFP1-B0015 with XbaI, PstI (using 10×M buffer)
Ligation
- Thu Jun 13
Ligate K1084007 with B0034-mRFP1-B0015
Transformation
- Thu Jun 13
K1084007-B0034-mRFP1-B0015
1µL DNA to DH5α
Colony PCR
- Mon Jun 17
K1084007-B0034-mRFP1-B0015
PCR
- Tue Jun 18 and Wed Jun 19
K1084007(colony)
ranori-f, 200dn, KOD FX Neo
We got random promter.

B0034-LacZα-B0015(colony)
100up, 200dn, KOD FX Neo
Digestion
- Thu Jun 20
Cut random promoter with PstI, SpeI (using 10×H buffer)
Cut B0034-LacZα-B0015 with XbaI, PstI (using 10×M buffer)
Ligation
- Thu Jun 20
Ligate random promoter with B0034-LacZα-B0015
Transformation
- Thu Jun 20
1µL DNA to JM109
to X-Gal plate
Culture
- Fri Jun 21
random promoter-B0034-LacZα-B0015
to X-Gal 50mg/mL, X-Gal 5mg/mL and LBC
We got some blue colonies with different levels of blue.
Culture
- Sat Jun 22
random promoter-B0034-LacZα-B0015 ×10 samples
random promoter-B0034-mRFP1-B0015 ×9 samples
K1084007-B0034-mRFP1-B0015
Mini-prep
- Sun Jun 23
random promoter-B0034-LacZα-B0015 ×10 samples
random promoter-B0034-mRFP1-B0015 ×9 samples
K1084007-B0034-mRFP1-B0015
Culture
- Wed Jun 26
random promoter-B0034-mRFP1-B0015
K1084007-B0034-mRFP1-B0015
random promoter-B0034-mRFP1-B0015 ×20 samples
PCR
- Thu Jun 27
random promoter-B0034-mRFP1-B0015 ×2 samples
random promoter-B0034-LacZα-B0015 ×2 samples
100up, 200dn, KOD FX Neo
PCR
- Fri Jun 28
J23112, J23103, J23117, J23106, J23111, J23100
100up, 200dn, KOD FX Neo
Digestion
- Fri Jun 28
Cut (J23112, J23103, J23117, J23106, J23111, J23100) with EcoRI, PstI (using 10×H buffer)
Cut B0034-mRFP1-B0015 with EcoRI, PstI (using 10×H buffer)
Antarctic phophatase
- Fri Jun 28
B0034-mRFP1-B0015
Ligation
- Sat Jun 29
Ligate (J23112, J23103, J23117, J23106, J23111, J23100) with B0034-mRFP1-B0015
Transformation
- Sat Jun 29 and Fri Jul 5
(J23112, J23103, J23117, J23106, J23111, J23100)-B0034-mRFP1-B0015
1µL DNA to JM109
pre culture
Culture
- Mon Jul 1
(J23112, J23103, J23117, J23106, J23111, J23100) with B0034-mRFP1-B0015
We picked up some colonies to a X-Gal LBC plate. We confirmed that colonies were white.

Transformation

Sat Jul 6

R0040-B0034-mRFP1-B0015

Culture
- Mon Jul 8
(J23112, J23103, J23117, J23106, J23111, J23100, R0040, R0010)-B0034-mRFP1-B0015
Transformation
- Thu Jul 11
random promoter-B0034-mRFP1-B0015
1µL DNA to JM109 to X-Gal plate
PCR & PCR purification
- Mon Jul 15
J23112, J23103, J23117(colony)
100up, 200dn, KOD FX Neo
Digestion
- Mon Jul 15
Cut J23112, J23103, J23117 with EcoRI, PstI (using 10×H buffer)
Cut random promoter-B0034-LacZα-B0015 on pSB1C3 with EcoRI, PstI (using 10×M buffer)
Antarctic phosphatase
- Tue Jul 16
pSB1C3
Ligation
- Tue Jul 16
Ligate (J23112, J23103, J23117) with pSB1C3
Transformation
- Tue Jul 16 and Fri Jul 19
1µL DNA to JM109 to 10 X-Gal plates
Culture
- Fri Jul 26
We cultured some colonies from Jul 19 and 23 plates to X-Gal plate.
Assay
- Wed Aug 7
Assay for RFP expression
Culture
- Thu Aug 8, Fri Aug 9 and Sat Aug 10
random promoter-B0034-mRFP1-B0015
Mini-prep
- Sat Aug 10
random promoter-B0034-mRFP1-B0015(cultured Aug 8)
Complete!

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      <div id="hokkaidou-contents">
 <!-- end header / begin contents -->
-<h2></h2>
+<h2>RandomPromtoer-B0034-mRFP1-B0015</h2>
 <div class="workflow-container">
    <ul class="steps">
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        <div class="step-tile"><h2>Start</h2></div>
      </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Tue Jun 11</time> </li> </ul> <div class="step-recipe"> B0034-mRFP1-B0015<br> 100up, 200dn, KOD Plus Neo </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Get K1084007 from our stock</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Tue Jun 11</time> </li> </ul> <div class="step-recipe"> </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">PCR purification</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed Jun 12</time> </li> </ul> <div class="step-recipe"> B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Digestion</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed Jun 12</time> </li> </ul> <div class="step-recipe"> Cut K1084007 with SpeI, PstI (using 10&times;H buffer)<br> Cut B0034-mRFP1-B0015 with XbaI, PstI (using 10&times;M buffer) </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Ligation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Jun 13</time> </li> </ul> <div class="step-recipe"> Ligate K1084007 with B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Jun 13</time> </li> </ul> <div class="step-recipe"> K1084007-B0034-mRFP1-B0015<br> 1&micro;L DNA to DH5&alpha; </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Colony PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jun 17</time> </li> </ul> <div class="step-recipe"> K1084007-B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Tue Jun 18 and Wed Jun 19</time> </li> </ul> <div class="step-recipe"> K1084007(colony)<br> ranori-f, 200dn, KOD FX Neo<br> We got random promter.<br><br> B0034-LacZ&alpha;-B0015(colony)<br> 100up, 200dn, KOD FX Neo </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Digestion</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Jun 20</time> </li> </ul> <div class="step-recipe"> Cut random promoter with PstI, SpeI (using 10&times;H buffer)<br> Cut B0034-LacZ&alpha;-B0015 with XbaI, PstI (using 10&times;M buffer) </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Ligation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Jun 20</time> </li> </ul> <div class="step-recipe"> Ligate random promoter with B0034-LacZ&alpha;-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Jun 20</time> </li> </ul> <div class="step-recipe"> 1&micro;L DNA to JM109<br> to X-Gal plate </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Jun 21</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-LacZ&alpha;-B0015<br> to X-Gal 50mg/mL, X-Gal 5mg/mL and LBC<br> We got some blue colonies with different levels of blue. </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat Jun 22</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-LacZ&alpha;-B0015 &times;10 samples<br> random promoter-B0034-mRFP1-B0015 &times;9 samples<br> K1084007-B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Mini-prep</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun Jun 23</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-LacZ&alpha;-B0015 &times;10 samples<br> random promoter-B0034-mRFP1-B0015 &times;9 samples<br> K1084007-B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed Jun 26</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015<br> K1084007-B0034-mRFP1-B0015<br> random promoter-B0034-mRFP1-B0015 &times;20 samples </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Jun 27</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015 &times;2 samples<br> random promoter-B0034-LacZ&alpha;-B0015 &times;2 samples<br> 100up, 200dn, KOD FX Neo </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Jun 28</time> </li> </ul> <div class="step-recipe"> J23112, J23103, J23117, J23106, J23111, J23100<br> 100up, 200dn, KOD FX Neo </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Digestion</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Jun 28</time> </li> </ul> <div class="step-recipe"> Cut (J23112, J23103, J23117, J23106, J23111, J23100) with EcoRI, PstI (using 10&times;H buffer)<br> Cut B0034-mRFP1-B0015 with EcoRI, PstI (using 10&times;H buffer) </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Antarctic phophatase</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Jun 28</time> </li> </ul> <div class="step-recipe"> B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Ligation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat Jun 29</time> </li> </ul> <div class="step-recipe"> Ligate (J23112, J23103, J23117, J23106, J23111, J23100) with B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat Jun 29 and Fri Jul 5</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111, J23100)-B0034-mRFP1-B0015<br> 1&micro;L DNA to JM109<br> pre culture </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jul 1</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111, J23100) with B0034-mRFP1-B0015<br> We picked up some colonies to a X-Gal LBC plate. We confirmed that colonies were white. </div> </div> </div> </li
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat Jul 6</time> </li> </ul> <div class="step-recipe"> R0040-B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jul 8</time> </li> </ul> <div class="step-recipe"> (J23112, J23103, J23117, J23106, J23111, J23100, R0040, R0010)-B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Jul 11</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015<br> 1&micro;L DNA to JM109 to X-Gal plate </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">PCR & PCR purification</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jul 15</time> </li> </ul> <div class="step-recipe"> J23112, J23103, J23117(colony)<br> 100up, 200dn, KOD FX Neo </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Digestion</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jul 15</time> </li> </ul> <div class="step-recipe"> Cut J23112, J23103, J23117 with EcoRI, PstI (using 10&times;H buffer)<br> Cut random promoter-B0034-LacZ&alpha;-B0015 on pSB1C3 with EcoRI, PstI (using 10&times;M buffer) </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Antarctic phosphatase</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Tue Jul 16</time> </li> </ul> <div class="step-recipe"> pSB1C3 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Ligation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Tue Jul 16</time> </li> </ul> <div class="step-recipe"> Ligate (J23112, J23103, J23117) with pSB1C3 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Tue Jul 16 and Fri Jul 19</time> </li> </ul> <div class="step-recipe"> 1&micro;L DNA to JM109 to 10 X-Gal plates </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Jul 26</time> </li> </ul> <div class="step-recipe"> We cultured some colonies from Jul 19 and 23 plates to X-Gal plate. </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Assay</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed Aug 7</time> </li> </ul> <div class="step-recipe"> Assay for RFP expression </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Aug 8, Fri Aug 9 and Sat Aug 10</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015 </div> </div> </div> </li>
+<li class="step"> <div class="step-tile"> <h2 class="step-title">Mini-prep</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat Aug 10</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-mRFP1-B0015(cultured Aug 8) </div> </div> </div> </li>
      <li class="step complete">
        <div class="step-tile"><h2>Complete!</h2></div>

Team:HokkaidoU Japan/Notebook/Promoter RandomPromtoer-B0034-mRFP1-B0015

From 2013.igem.org

Revision as of 06:55, 27 September 2013

HOKKAIDO U

Maestro E.coli

Notebook

RandomPromtoer-B0034-mRFP1-B0015

Start

PCR

Get K1084007 from our stock

PCR purification

Digestion

Ligation

Transformation

Colony PCR

PCR

Digestion

Ligation

Transformation

Culture

Culture

Mini-prep

Culture

PCR

PCR

Digestion

Antarctic phophatase

Ligation

Transformation

Culture

Transformation

Culture

Transformation

PCR & PCR purification

Digestion

Antarctic phosphatase

Ligation

Transformation

Culture

Assay

Culture

Mini-prep

Complete!