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-lacI 5
SacI 0.2×22
XbaI 0.2
10×M Buffer 2
ddH2O 12.6

Result: false positive transformation




PCR system
Volume (μL)
10×PCR buffer 2 ×22
transtart 0.1
dNTP 2
template 2
F-Primer 0.4
R-Primer 0.4
ddH2O 13.1


conditions
Temperature(°C) Time Length(0:00:00)
94 02:00
94 00:20 30 cycles
60 00:30 30 cycles
72 07:00


Reinforce the result of false positive transformation




Colony PCR


Volume (μL)
Buffer 1.5 ×20
Easy tag 0.1
dNTP 1.5
primer-R 0.5
primer-F 0.5
ddH2O 11