The PCR reaction for further sequencing analysis consisted of 1 unit of ExTaq polymerase (TaKaRa) in a total reaction of 50 µL.
All amplifications were performed for a total of 35 cycles of 1 min at 95°C,30sat55°C, and 1 min at 72°C.
For STS markers developed by Ohtsubo et al. (13, 17) and Ohtsubo and Nakamura (12), the initial denaturation was at 95°C for 5 min, followed by 40 cycles of 96°C for 1 min, 62 °C for 1 min, and 72°C for 2 min.