A | B | |
Component | Volume (uL) | |
5x green buffer | 5 | |
dNTPs | 0.5 | |
Forward primer | 1.25 | |
Reverse primer | 1.25 | |
Template | 1 | |
GoTaq | 0.125 | |
MgCl2 | 1 | |
H2O | 14.875 | |
Total | 25 |
A | B | C | |
Step | Temperature (deg C) | Time | |
Initial Denaturation | 95 | 2 min | |
30 cycles | 95 | 30 s | |
Ta | 60 s | ||
72 | 1 min per kb | ||
Final Extension | 72 | 10 min | |
Hold | 4 | HOLD |