Template:Kyoto/Notebook/Sep 11

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Contents

Sep 11

PCR

Tatsui

pKaiBC(200p)10*buffer KOD plusdNTPsMgSO4primer mix(F&R)KOD plusMilliQtotal
0.52.52.51.51.50.516 25
PreDenatureDenatureAnnealingExtensioncycle
 94°C 98°C 60°C 68°C30
2min 10sec 30sec 30sec 
pKaiBC(400p)10*buffer KOD plusdNTPsMgSO4primer mix(F&R)KOD plusMilliQtotal
12.52.51.51.50.515.5 25
PreDenatureDenatureAnnealingExtensioncycle
 94°C 98°C 60°C 68°C30
2min 10sec 30sec 30sec 
pKaiBC(800p)10*buffer KOD plusdNTPsMgSO4primer mix(F&R)KOD plusMilliQtotal
22.52.51.51.50.514.5 25
PreDenatureDenatureAnnealingExtensioncycle
 94°C 98°C 60°C 68°C30
2min 10sec 30sec 30sec 
pKaiBC(1ng)10*buffer KOD plusdNTPsMgSO4primer mix(F&R)KOD plusMilliQtotal
2.52.52.51.51.50.514 25
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
2 cuts(°°) µL µL µL µL µL µL µL
NC µL µL µL µL µL µL µL