PCR condition
PCR amplification was done in a total volume of
30 μl. Each reaction consisted of 1X master mixed
with 10 pM of each primer per reactions and 10 ng
of template plasmid. PCR was performed under the
following conditions: Initial denaturation at 95°C
for 4 min followed by 35 cycles of at 95°C for 30 sec,
annealing at 54°C for 30 sec, extension at 72°C for
80 sec, and final extension at 72°C for 10 min.