梨品种ISSR-PCR反应体系的优化与应用

Optimization and application of ISSR-PCR reaction system in pear cultivars

摘要: 以海子梨渊Haizili冤品种的DNA 为材料袁对影响ISSR-PCR 扩增的重要因子进行单因素的优化设计袁建立了最佳体系: 25 滋L 反应体系中含2.5 滋L 10伊buffer尧0.75 U Taq DNA 聚合酶尧0.2 mmol/L dNTPs尧2.5 mmol/LMgCl2尧0.25 滋mol/L 引物尧10~60 ng 模板DNA遥将最佳体系应用于黔西南的22 个梨品种袁证实了该体系的重复性好袁很稳定遥

Abstract: The influencing factors in ISSR reaction system were studied by using genomic DNA of different pear cultivars. ISSR-PCR optimized reaction systems were eatablished for pear cultivars. The total volume of 25 滋L included 2.5 滋L10伊buffer, 2.5 mmol/L MgCl2, 0.2 mmol/L dNTPs, 0.25 滋mol/L primer, 10~60 ng template DNA and 0.75 U unit of DNA polymerase. 22 different pear cultivars were used to check the stabilization of the optimized reaction system. The results showed that the optimal reaction system of ISSR-PCR of pear was stable, repeatable and effective .