Leaf tissues were collected from transgenics and wild type plants grown in glass jars and genomic DNA was isolated by DNeasy Plant Mini kit (Qiagen). Genomic DNA was digested with BglII and BamHI and run on 1% agarose gel–electrophoresis, denatured and blotted on Hybond positive nylon membrane as described (Kumar and Daniell, 2004). The probe was prepared according to AlkPhos Direct Labelling Reagents kit (GE Healthcare) using nptII gene (amplified with primers, nptIIF and nptIIR) and Southern hybridization was carried out according to AlkPhos kit (GE Healthcare) instructions ( Fig. 5).