IGEM:Peking University/2008/Notebook/Group 3/2008/07/29

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PKU_080729_G3_1

Enzyme cutting-testing-pGREG503-lacI

System1:Giving up cause no EcoRI


System2:

Volume (μL)
pGREG503-lacI5
SacI0.2×22
XbaI0.2
10×M Buffer2
ddH2O12.6

Result: false positive transformation




PCR system
Volume (μL)
10×PCR buffer2×22
transtart0.1
dNTP2
template2
F-Primer0.4
R-Primer0.4
ddH2O13.1


conditions
Temperature(°C)Time Length(0:00:00)
9402:00
9400:2030 cycles
6000:3030 cycles
7207:00


Reinforce the result of false positive transformation




Colony PCR


Volume (μL)
Buffer 1.5×20
Easy tag0.1
dNTP1.5
primer-R0.5
primer-F0.5
ddH2O11





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