The presence of transgene in putative transformed shoots was
confirmed through PCR amplification of cry1Aa3 and nptII genes
using gene-specific primers. Genomic DNAwas isolated from leaves
of putative transformed shoots and non-transgenic (WT) plant by
the Purelink plant DNA purification kit (Invitrogen, CA, USA). The
primer pair for amplification of cry1Aa3 gene was F-50-tggctaacaacccaaacatcaacg-
30 and R-50-tgaaactaccatcgaagttctcga-30 , which
was expected to produce an amplified product of 840 bp. The expected
amplification product size for nptII gene-specific primer pair
(F-50-gcgataccgtaaagcacgaggaa-30 and R-50-gatggattgcacgcaggttctc-