Confirmation of transgenic lines by polymerase chain reaction
(PCR) and Southern blot analysis
Genomic DNA was isolated by c etyl t rimethyla mmonium b romide (CTAB) method (Nickrent, 1994) from
4-week old plants of the T1 generation. The presence of trans-gene in
the plants was confirmed by PCR using 35S forward internal primer andFbβCA3 reverse primer to ensure the incorporation of the whole
cassette in sense orientation. The presence of FbβCA3 transgene
was analysed by Southern blot analysis, as described by Kandoi et al.
(2016).