Template:Kyoto/Notebook/Sep 11

From 2013.igem.org

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(→Restriction Enzyme Digestion)
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|pSB4K5(E+S) negative||0.5µL|| 0µL|| 0µL|| 0µL|| 0µL|| 1µL|| 1µL|| 7.5µL|| 10µL
|pSB4K5(E+S) negative||0.5µL|| 0µL|| 0µL|| 0µL|| 0µL|| 1µL|| 1µL|| 7.5µL|| 10µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL
+
|RBS-lacZα-DT(E+S)|| 10µL|| 1µL|| 0µL|| 1µL|| 0µL|| 3µL|| 3µL|| 12µL|| 30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL
+
|RBS-lacZα-DT(E+S) negative|| 0.5µL|| 0µL|| 0µL|| 0µL|| 0µL|| 1µL|| 1µL|| 7.5µL|| 10µL
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-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL
+
|Ptet-pT181 antisense(S+P)|| 10.8µL|| 0µL|| 0µL|| 1µL|| 1µL|| 3µL|| 3µL|| 12µL|| 30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL
+
|Ptet-pT181 antisense(S+P) negative|| 0.5µL|| 0µL|| 0µL|| 0µL|| 0µL|| 1µL|| 1µL|| 7.5µL|| 10µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL
+
|Pcon-RBS-lacZα-DT(E+S)|| 7.1µL|| 1µL|| 0µL|| 1µL|| 0µL|| 3µL|| 3µL|| 14.9µL|| 30µL
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|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL
+
|Pcon-RBS-lacZα-DT(E+S) negative|| 0.4µL|| 0µL|| 0µL|| 0µL|| 0µL|| 1µL|| 1µL|| 7.6µL|| 10µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL
+
|RBS-lysis2-DT(X+P)|| 22µL|| 0µL|| 1µL|| 0µL|| 1µL|| 3µL|| 3µL|| 0µL|| 30µL
|-
|-
-
|NC|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL|| µL
+
|RBS-lysis2-DT(X+P) negative|| 1.4µL|| 0µL|| 0µL|| 0µL|| 0µL|| 1µL|| 1µL|| 6.6µL|| 10µL
|}
|}
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Revision as of 13:04, 24 September 2013

Contents

Sep 11

PCR

Tatsui

pKaiBC(200p)10*buffer KOD plusdNTPsMgSO4primer mix(F&R)KOD plusMilliQtotal
0.52.52.51.51.50.51625
PreDenatureDenatureAnnealingExtensioncycle
　94°C　98°C　60°C　68°C30
2min　10sec　30sec　30sec　
pKaiBC(400p)10*buffer KOD plusdNTPsMgSO4primer mix(F&R)KOD plusMilliQtotal
12.52.51.51.50.515.525
PreDenatureDenatureAnnealingExtensioncycle
　94°C　98°C　60°C　68°C30
2min　10sec　30sec　30sec　
pKaiBC(800p)10*buffer KOD plusdNTPsMgSO4primer mix(F&R)KOD plusMilliQtotal
22.52.51.51.50.514.525
PreDenatureDenatureAnnealingExtensioncycle
　94°C　98°C　60°C　68°C30
2min　10sec　30sec　30sec　
pKaiBC(1ng)10*buffer KOD plusdNTPsMgSO4primer mix(F&R)KOD plusMilliQtotal
2.52.52.51.51.50.51425
PreDenatureDenatureAnnealingExtensioncycle
　94°C　98°C　60°C　68°C30
2min　10sec　30sec　30sec　

colony PCR

Tatsui

Samplebase pair
9/9Pcon-lacZα(p3B4k5)712bp
9/9Pcon-spinach-DT(p3B4k5)605bp
PreDenatureDenatureAnnealingExtensioncycle
　94°C　94°C　55°C　68°C--
5min30s　30s　42s　
Samplebase pair
Pbad\araC-RBS-luxI-DT
PreDenatureDenatureAnnealingExtensioncycle
　94°C　94°C　55°C　68°C--
5min30s　30s　2min　
Samplebase pair
9/9Pcon+RBS-luxI-DT1079bp
9/10Pcon+sptamer12_1R-DT584bp
9/10DT+Pcon-pT181 attenuater 1781bp
9/10Fusion1 attenuater(p3B1C3)-1638bp
PreDenatureDenatureAnnealingExtensioncycle
　94°C　94°C　55°C　68°C--
50sec　30sec　30sec　1min　

Restriction Enzyme Digestion

Nakamoto

DNAEXSPbufferBSAMilliQTotal
pSB4K5(E+S)9.4µL 1µL 0µL 1µL 0µL 3µL 3µL 12.6µL30 µL
pSB4K5(E+S) negative0.5µL 0µL 0µL 0µL 0µL 1µL 1µL 7.5µL 10µL
RBS-lacZα-DT(E+S) 10µL 1µL 0µL 1µL 0µL 3µL 3µL 12µL 30µL
RBS-lacZα-DT(E+S) negative 0.5µL 0µL 0µL 0µL 0µL 1µL 1µL 7.5µL 10µL
Ptet-pT181 antisense(S+P) 10.8µL 0µL 0µL 1µL 1µL 3µL 3µL 12µL 30µL
Ptet-pT181 antisense(S+P) negative 0.5µL 0µL 0µL 0µL 0µL 1µL 1µL 7.5µL 10µL
Pcon-RBS-lacZα-DT(E+S) 7.1µL 1µL 0µL 1µL 0µL 3µL 3µL 14.9µL 30µL
Pcon-RBS-lacZα-DT(E+S) negative 0.4µL 0µL 0µL 0µL 0µL 1µL 1µL 7.6µL 10µL
RBS-lysis2-DT(X+P) 22µL 0µL 1µL 0µL 1µL 3µL 3µL 0µL 30µL
RBS-lysis2-DT(X+P) negative 1.4µL 0µL 0µL 0µL 0µL 1µL 1µL 6.6µL 10µL
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