10 pmol of forward or reverse primer, 2.3 µL of
purified PCR product, and sterile distilled water to
a final volume of 10 µL for each sample. For cycle
sequencing, a Perkin Elmer 9600 thermal cycler
was used under the following conditions: 25 cycles
at 96°C for 30 seconds, 50°C for 15 seconds, and
60°C for 4 minutes. After sequencing, each sample was added to a sephadex-G-50 column (spin
50-mini-column; BioMax Inc., Odenton, MD,
USA) and centrifuged at 1500g for 3 minutes.