桃SSR反应体系的优化

Optimization of SSR reaction system of peach

以‘新大久保’桃为试材,利用正交设计直观分析法和2因素完全随机试验优化了桃SSR技术中PCR反应体系.试验结果表明,适宜桃遗传分析的SSR技术体系为:在25 μL反应体系中Taq DNA聚合酶和DNA最适用量分别为1 U和50～100 ng;Mg2+ 、dNTP和引物最适终浓度分别为1.5～2.0 mmol/L、0.20～0.28 mmol/L和0.2～0.6 μmol/L.利用30个桃品种验证此反应体系,变性聚丙烯酰胺凝胶电泳检测结果显示,扩增产物在100～300 bp,多态性高,且反应体系的稳定性和可重复性好.

‘Shinokubo＇ peach （Prunus persica （L.） Batsch. ） was used to set up a SSR-PCR amplification system for peach. The results showed that the optimal SSR-PCR reaction system for peach was 1.5-2.0 mmol/L Mg^2＋ concentration, 0.20-0.28 mmol/L dNTP, 0.2-0.6 ,umol/L primer, 50-100 ng DNA and 1 U in 25 μL Taq DNA polymerase. To retest the above SSR system, 30 cultivars of peach were used. SSR fragments between 100-300 bp were detected by polyacrylamide gel （8%）, indicating that the SSR reaction system was steady and reproducible.