The modified Dellaporta et al. (1983) DNA extraction
procedure was used to extract DNA from the plant
materials (oven-dried young leaf samples) of the
parents and their progeny. The DNA was stored at
-20 C until when needed for genotyping. The DNA
samples of the two parents of each population were
amplified with each of the 542 simple sequence repeat
(SSR) markers (represented by a pair of primers—
forward and reverse) using the protocols by Williams
et al. (1990). The PCR products (about 15 ll each)
were denatured at 95 C and 3 ll of the denatured
products were electrophoresed on 4 %polyacrylamide
gels using Bio-Rad sequencing apparatus (Bio-Rad
Inc., USA) for 15–40 min depending on the size (bp)
of the marker. The gel was then developed, air-dried
and scanned. Patterns were observed for banding
differences between the EB and LB parents and primer
pairs that distinguished between the parents were
selected as polymorphic markers (pm-p).