Figure 3
Molecular analysis of marker gene excision. A Multiplex PCR analysis of F1 hybrid line LA2CC3. Lane '+’, mix of pBKhgASAL and pBK16.2 plasmids as positive control; lane '- ’ genomic DNA of an untransformed plant. Lanes 1–30, 30 F1 hybrid progenies of line LA2CC3. The 362 bp and 1.1 kb amplicon derives from the ASAL and cre genes respectively and B PCR with hpt specific primers of 16 ASAL + cre + plants of F1 hybrid line LA2CC3. Lanes 1–16, F1 hybrid plants, where plant no. 3, 6, 9, 12, 14, 18, 23, 24 and 27 showed absence of hpt. Lane ' + ', pBKhgASAL ; lane ' – ', genomic DNA of an untransformed plant. The 980 bp amplicon derives from the hpt gene. Lane M in A and B, DNA molecular weight marker (GenerulerTM, MBI Fermentas, UK).