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
|
|