PCR analysis of transformed plants
Genomic DNA was extracted from young
leaves of transformed rice plants and untransformed
control plants using modified CTAB method (Hwang
and Kim, 2000). The extracted DNA was subjected
to PCR methods using primers specific to OSB2
gene. The primers were OSB2cdsF: 5'-ATGGCATCT
GCTCCTCCAGTTCAGG-3' and OSB2cdsR: 5'-TTA
CGGCGCCTTCCCCTGTCC-3'. The PCR amplification
was carried out in a 20 l reaction mixture
containing GoTaq® Green Master Mix (Promega,
USA), 100 ng of DNA and 0.5 M of each primer.
The PCR profile was initially at 95 oC for 3 min,
followed by 35 cycles at 95 oC for 1 min, 68 oC for 1
min and 72 oC for 1.5 min and 5 min at 72 oC for
final extension. The expected PCR products of 1,356
bp were analyzed on a 1 % (w/v) agarose gel by
electrophoresis.