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.
All amplifications using primers reported by Bao et al. (2, 3) were performed under the following conditions: 5 min at 94°C followed by 35 cycles of 45 s at 94°C, 1 min at 55°C, and 1 min at 72°C, with a final extension of
7 min at 72°C.
Amplified PCR products were analyzed by electrophoresis on 3% agarose gels stained with ethidium bromide and/or by nondenaturing electrophoresis on 8% polyacrylamide gels stained with ethidium bromide (model MGV, CBS Scientific Co.).