1 µl DNA (or ddH2O for blank sample). If you are performing a colony PCR, pick up the desired colony from a plate with a tip and dip it in the solution.
0.2 µl Taq Polymerase
250 nM VF2 primer
250 nM VR primer
A proper amount of ddH2O to have 20 µl of total reaction volume
into an eppendorf tube.
Put the eppendorf tube in the thermal cycler and set this program:
95°C 10 min
CYCLE:
95°C 30 sec
60°C 1 min
72°C 1-3 min
for 35 cycles
72°C 7 min
16°C forever.
Now you can add a loading buffer to the solution and perform electrophoresis to check the amplified sequence length.
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