Figure 2. Confirmation of transgenic nature of hairy root clones by PCR (a) and Southern analysis (b). Lanes 1-10; LMG-150; A4 (1); A4 (2); A4 (3); A 2/83(1); A 2/83(2); A 2/83(3); A 20/83(1); A 20/83(2); A 20/83(3); M = Marker i.e., 100 bp ladder; C = untransformed control from seedling explants. PCR was performed using rolA gene specific primer sets (forward- 5' AGA ATG GAA TTA GCC GGA CTA 3' and reverse- 5' GTA TTA ATC CCG TAG GTT TGT TT -3') (Sigma, USA), which were designed using Primer3 software. The amplified DNA was run on 0.8% (w/v) agarose gel to separate the DNA fragments, transferred to nylon membrane (BioBond^TM-Plus, Sigma). Southern analysis (b) was done by probing with a psoralen biotin (Ambion Inc, USA) labelled 308 bp fragment of the rolA gene.