Molecular confirmation and expression analysis of cry2Aa in transgenic brinjal plants. PCR amplification of the cry2Aa (A) and nptII gene (B) from genomic DNA of putative transgenic plants (T0). Lanes: M1, 1-kb DNA ladder; W, water control; C, genomic DNA from wildtype plant; 1-16, putative transgenic plants. (C) Southern blot analysis of transgenic plants. Lane 1, 3, 4, 5, 6 and 7 shows single copy integration, Lane 2 shows triple copy integration while lane 8 shows double copy integration M: λ DNA/HindIII ladder (Thermo Scientific). The left border (LB) junction fragment of the T-DNA is greater than 2.6 kb in pBinAR-UBI:Cry2Aa binary vector (the distance between HindIII and LB). (D) The qRT-PCR analysis shows relative fold change in gene expression of cry2Aa transcript in T2 generation of transgenic brinjal plants. The expression of cry2Aa was determined by using 18S rRNA as internal control. E1, E3, E4 and E7 represent T2 generation transgenic lines.