Template:Kyoto/Notebook/Sep 11

From 2013.igem.org

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|pSB4K5(E+S)||9.4µL|| 1µL|| 0µL|| 1µL|| 0µL|| 3µL|| 3µL|| 12.6µL||30 µL
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|pSB4K5(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
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Revision as of 12:45, 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
NC µL µL µL µL µL µL µL µL µL
NC µL µL µL µL µL µL µL µL µL
NC µL µL µL µL µL µL µL µL µL
NC µL µL µL µL µL µL µL µL µL
NC µL µL µL µL µL µL µL µL µL
NC µL µL µL µL µL µL µL µL µL
NC µL µL µL µL µL µL µL µL µL
NC µL µL µL µL µL µL µL µL µL
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