Template:Kyoto/Notebook/Aug 29

From 2013.igem.org

(Difference between revisions)
(→Liquid Culture)
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Line 17: Line 17:
|5min||30s||30s||30s||30cycles
|5min||30s||30s||30s||30cycles
|}
|}
-
[[File:igku_Aug19electrophoresis1.png]]
+
 
 +
{| class="wikitable"
 +
!Lane||Sample
 +
|-
 +
|1||100bp ladder
 +
|-
 +
|2||8/27 Pcon-pT181antisense-1
 +
|-
 +
|3||8/26 pT181attenuator-DT-1
 +
|-
 +
|4||100bp ladder
 +
|}
 +
[[File:Igku Aug29 Electrophoresis(ColoP)(N1)pic.jpg]]<br>
</div>
</div>
Line 44: Line 56:
|9||8/21 RBS-lysis2-14(Colony PCR prodution)
|9||8/21 RBS-lysis2-14(Colony PCR prodution)
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:Igku Aug29 Electrophoresis1(N②).jpg]]<br>
</div>
</div>
-
===miniprep===
+
 
 +
===Miniprep===
<div class="experiment">
<div class="experiment">
<span class="author">No Name</span>
<span class="author">No Name</span>
Line 54: Line 67:
|Plac||153.7||2.03||2.10
|Plac||153.7||2.03||2.10
|-
|-
-
|Pλ-luxⅠ-(1)||76.5||2.07||2.06
+
|P&lambda;-luxⅠ-(1)||76.5||2.07||2.06
|-
|-
-
|Pλ-luxⅠ-(2)||86.4||2.08||2.10
+
|P&lambda;-luxⅠ-(2)||86.4||2.08||2.10
|-
|-
|DT||166.2||1.65||2.12
|DT||166.2||1.65||2.12
Line 74: Line 87:
|8/21 RBS-Lysis2-12||Plusgrow medium(+Amp)
|8/21 RBS-Lysis2-12||Plusgrow medium(+Amp)
|-
|-
-
|8/21 RBS-Lysis2-14||Plusgrow medium(+Amp)　
+
|8/21 RBS-Lysis2-14||Plusgrow medium(+Amp)
|}
|}
</div>
</div>
===Restriction Enzyme Digestion===
===Restriction Enzyme Digestion===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
! ||8/22 psB1C3-(2)||EcoRI||SpeI||XbaI||PstI||BufferB||BufferD||BSA||MilliQ||total
 +
|-
 +
|2 cuts||8&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||0&micro;L||3&micro;L||0&micro;L||0.3&micro;L||17.7&micro;L||30&micro;L
 +
|-
 +
|NC||1&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0&micro;L||0.1&micro;L||7.9&micro;L||10&micro;L
 +
|-
 +
|2 cuts||8&micro;L||0&micro;L||0&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||3&micro;L||0.3&micro;L||17.7&micro;L||30&micro;L
 +
|-
 +
|NC||1&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||7.9&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 tRMA-spinach(1)||XbaI||SpeI||BufferB||BSA||MilliQ||total
 +
|-
 +
|2 cuts||4&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||21.7&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.4&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 tetR aptamer12-1R(1)||EcoRI||SpeI||BufferB||BSA||MilliQ||total
 +
|-
 +
|2 cuts||3&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||22.7&micro;L||30&micro;L
 +
|-
 +
|NC||0.4&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.5&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 PT181 attenuator-(2)||EcoRI||SpeI||XbaI||PstI||BufferB||BufferD||BSA||MilliQ||total
 +
|-
 +
|2 cuts||4&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||0&micro;L||3&micro;L||0&micro;L||0.3&micro;L||21.7&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0&micro;L||0.1&micro;L||8.4&micro;L||10&micro;L
 +
|-
 +
|2 cuts||4&micro;L||0&micro;L||0&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||3&micro;L||0.3&micro;L||21.7&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.4&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 PT181 antisense-(2)||EcoRI||SpeI||XbaI||PstI||BufferB||BufferD||BSA||MilliQ||total
 +
|-
 +
|2 cuts||10&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||0&micro;L||3&micro;L||0&micro;L||0.3&micro;L||15.7&micro;L||30&micro;L
 +
|-
 +
|NC||1&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0&micro;L||0.1&micro;L||7.9&micro;L||10&micro;L
 +
|-
 +
|2 cuts||10&micro;L||0&micro;L||0&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||3&micro;L||0.3&micro;L||15.7&micro;L||30&micro;L
 +
|-
 +
|NC||1&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||7.9&micro;L||10&micro;L
 +
|}
 +
*incubated 37&deg;C 1hour
 +
[[File:Igku Aug29 Restriction Enzyme Digestion(N③).jpg]]<br>
 +
</div>
 +
===Transformation===
===Transformation===
-
===M9 Medium Plate===
+
<div class="experiment">
 +
<span class="author">No Name</span>
 +
{| class="wikitable"
 +
!Name||Sample||Competent Cells||Total||Plate
 +
|-
 +
|8/27 pT181 attenuator-DT||2&micro;L||20&micro;L||22&micro;L||CP
 +
|-
 +
|8/27 RBS-lysis3-DT||2&micro;L||20&micro;L||22&micro;L||CP
 +
|-
 +
|8/28 RBS-lysis3-DT||2&micro;L||20&micro;L||22&micro;L||CP
 +
|-
 +
|8/28 Pcon-RBS-GFP-DT-Pcon-RBS-LuxR-DT||2&micro;L||20&micro;L||22&micro;L||Amp
 +
|-
 +
|8/28 RBS-lysis1-DT||2&micro;L||20&micro;L||22&micro;L||CP
 +
|-
 +
|8/28 Plux-RBS-GFP-DT||2&micro;L||20&micro;L||22&micro;L||CP
 +
|-
 +
|8/28 Spinach-DT||2&micro;L||20&micro;L||22&micro;L||CP
 +
|}
 +
</div>
 +
 
 +
===Gel Extraction===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme1||Enzyme2
 +
|-
 +
|||100bp ladder||--||--
 +
|-
 +
|9||8/29 pT181attenuator(2)||EcoRI || SpeI
 +
|-
 +
|10||8/29 pT181attenuator(2)||EcoRI || SpeI
 +
|-
 +
|11||8/29 pT181attenuator(2)||XbaI||PstI
 +
|-
 +
|12||8/29 pT181attenuator(2)||XbaI||PstI
 +
|-
 +
|13||pT181antisense(2)||EcoRI||SpeI
 +
|-
 +
|14||pT181antisense(2)||EcoRI||SpeI
 +
|-
 +
|15||pT181antisense(2)||XbaI||PstI
 +
|-
 +
|16||pT181antisense(2)||XbaI||PstI
 +
|-
 +
|||100bp ladder||--||--
 +
|}
 +
[[File:Igku Aug29 Transformation1(N④-1).jpg]]<br>
 +
[[File:Igku Aug29 Transformation(N④-2).jpg]]<br>
 +
 
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|||100bp ladder||--||--
 +
|-
 +
|1||8/29 pSB1C3||EcoRI||SpeI
 +
|-
 +
|2||8/29 pSB1C3||EcoRI||SpeI
 +
|-
 +
|3||8/29 pSB1C3||XbaI||PstI
 +
|-
 +
|4||8/29 pSB1C3||XbaI||PstI
 +
|-
 +
|5||8/29 spinach||EcoRI||SpeI
 +
|-
 +
|6||8/29 spinach||EcoRI||SpeI
 +
|-
 +
|7||8/29 tetRaptarmer12-1R(1)||EcoRI||SpeI
 +
|-
 +
|8||8/29 tetRaptarmer12-1R(1)||EcoRI||SpeI
 +
|-
 +
|||100bp ladder||--||--
 +
|}
 +
 
 +
 
 +
[[File:Igku Aug29 Transformation(N⑤).jpg]]<br>
 +
[[File:Igku Aug29 Gel Extraction4-2.jpg]]<br>
 +
</div>

Latest revision as of 03:48, 28 September 2013

Contents

Aug 29

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles
LaneSample
1100bp ladder
28/27 Pcon-pT181antisense-1
38/26 pT181attenuator-DT-1
4100bp ladder

Igku Aug29 Electrophoresis(ColoP)(N1)pic.jpg

Electrophoresis

No name

LaneSample
1100bp ladder
28/21 RBS-lysis2-9(Colony PCR prodution)
38/21 RBS-lysis2-8(Colony PCR prodution)
48/21 RBS-lysis2-7(Colony PCR prodution)
58/21 RBS-lysis2-10(Colony PCR prodution)
68/21 RBS-lysis2-11(Colony PCR prodution)
78/21 RBS-lysis2-12(Colony PCR prodution)
88/21 RBS-lysis2-13(Colony PCR prodution)
98/21 RBS-lysis2-14(Colony PCR prodution)

Igku Aug29 Electrophoresis1(N②).jpg

Miniprep

No Name

DNAconcentration [µg/mL]260/280260/230
Plac153.72.032.10
Pλ-luxⅠ-(1)76.52.072.06
Pλ-luxⅠ-(2)86.42.082.10
DT166.21.652.12
J23100190.11.682.34

Liquid Culture

No name

Samplemedium
8/21 RBS-Lysis2-9Plusgrow medium(+Amp)
8/21 RBS-Lysis2-12Plusgrow medium(+Amp)
8/21 RBS-Lysis2-14Plusgrow medium(+Amp)

Restriction Enzyme Digestion

No name

8/22 psB1C3-(2)EcoRISpeIXbaIPstIBufferBBufferDBSAMilliQtotal
2 cuts8µL0.5µL0.5µL0µL0µL3µL0µL0.3µL17.7µL30µL
NC1µL0µL0µL0µL0µL1µL0µL0.1µL7.9µL10µL
2 cuts8µL0µL0µL0.5µL0.5µL0µL3µL0.3µL17.7µL30µL
NC1µL0µL0µL0µL0µL0µL1µL0.1µL7.9µL10µL
8/21 tRMA-spinach(1)XbaISpeIBufferBBSAMilliQtotal
2 cuts4µL0.5µL0.5µL3µL0.3µL21.7µL30µL
NC0.5µL0µL0µL1µL0.1µL8.4µL10µL
8/21 tetR aptamer12-1R(1)EcoRISpeIBufferBBSAMilliQtotal
2 cuts3µL0.5µL0.5µL3µL0.3µL22.7µL30µL
NC0.4µL0µL0µL1µL0.1µL8.5µL10µL
8/21 PT181 attenuator-(2)EcoRISpeIXbaIPstIBufferBBufferDBSAMilliQtotal
2 cuts4µL0.5µL0.5µL0µL0µL3µL0µL0.3µL21.7µL30µL
NC0.5µL0µL0µL0µL0µL1µL0µL0.1µL8.4µL10µL
2 cuts4µL0µL0µL0.5µL0.5µL0µL3µL0.3µL21.7µL30µL
NC0.5µL0µL0µL0µL0µL0µL1µL0.1µL8.4µL10µL
8/21 PT181 antisense-(2)EcoRISpeIXbaIPstIBufferBBufferDBSAMilliQtotal
2 cuts10µL0.5µL0.5µL0µL0µL3µL0µL0.3µL15.7µL30µL
NC1µL0µL0µL0µL0µL1µL0µL0.1µL7.9µL10µL
2 cuts10µL0µL0µL0.5µL0.5µL0µL3µL0.3µL15.7µL30µL
NC1µL0µL0µL0µL0µL0µL1µL0.1µL7.9µL10µL
  • incubated 37°C 1hour

Igku Aug29 Restriction Enzyme Digestion(N③).jpg

Transformation

No Name

NameSampleCompetent CellsTotalPlate
8/27 pT181 attenuator-DT2µL20µL22µLCP
8/27 RBS-lysis3-DT2µL20µL22µLCP
8/28 RBS-lysis3-DT2µL20µL22µLCP
8/28 Pcon-RBS-GFP-DT-Pcon-RBS-LuxR-DT2µL20µL22µLAmp
8/28 RBS-lysis1-DT2µL20µL22µLCP
8/28 Plux-RBS-GFP-DT2µL20µL22µLCP
8/28 Spinach-DT2µL20µL22µLCP

Gel Extraction

No name

LaneDNAEnzyme1Enzyme2
100bp ladder----
98/29 pT181attenuator(2)EcoRI SpeI
108/29 pT181attenuator(2)EcoRI SpeI
118/29 pT181attenuator(2)XbaIPstI
128/29 pT181attenuator(2)XbaIPstI
13pT181antisense(2)EcoRISpeI
14pT181antisense(2)EcoRISpeI
15pT181antisense(2)XbaIPstI
16pT181antisense(2)XbaIPstI
100bp ladder----

Igku Aug29 Transformation1(N④-1).jpg
Igku Aug29 Transformation(N④-2).jpg

LaneSampleEnzyme1Enzyme2
100bp ladder----
18/29 pSB1C3EcoRISpeI
28/29 pSB1C3EcoRISpeI
38/29 pSB1C3XbaIPstI
48/29 pSB1C3XbaIPstI
58/29 spinachEcoRISpeI
68/29 spinachEcoRISpeI
78/29 tetRaptarmer12-1R(1)EcoRISpeI
88/29 tetRaptarmer12-1R(1)EcoRISpeI
100bp ladder----


Igku Aug29 Transformation(N⑤).jpg
Igku Aug29 Gel Extraction4-2.jpg

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