梨属野生种质资源 SRAP －PCR 反应体系优化研究

摘要

采用正交直观分析法和新复极差法对影响梨属野生种质资源SRAP－PCR反应的5种因素（Mg2＋浓度、dNTPs浓度、引物浓度、Taq DNA聚合酶、模板DNA）4个水平进行优化筛选。结果表明，优化后的梨属SRAP－PCR反应体系为25μL，包括10×PCR buffer 2．5μL，2．0 mmol／L Mg2＋，200μmol／L dNTPs，0．4μmol／L引物，1．5 U Taq酶，60 ng模板DNA。%Taking Pyrus wild germplasms as materials, four levels of five factors influencing SRAP am-plification system were optimized using the orthogonal design-direct analysis and Duncan’s new multiple range tests.The five factors were as the concentrations of Mg2+, dNTPs, primers, Taq DNA polymerase and tem-plate DNA.The results showed that the optimized SRAP reaction system was 25 μL and contained 10 ×PCR buffer 2.5 μL, 2.0 mmol/L Mg2+, 200 μmol/L dNTPs, 0.4 μmol/L primers, 1.5 U Taq DNA polymerase and 60 ng template DNA.