甘西鼠尾草ISSR-PCR反应体系的建立与优化

Establishment and Optimization of Salvia przewalskii Maxim. ISSR-PCR Reaction System

采用单因素试验和4因素3水平的正交试验相结合的方法,建立和优化甘西鼠尾草稳定的ISSR-PCR反应体系。结果表明,甘西鼠尾草20 ul ISSR-PCR体系中各因素的最佳浓度:10×Buffer(Mg2+)2μl,d NTPs 0.4 mmol/L,Tag酶1.0 U,引物0.3μmol/L,DNA 30 ng。

Using the single factor test and the 4 factors and 3 level orthogonal test, the ISSR-PCR reaction system and amplification program for Salvia przewalskii Maxim were established. The study showed that the optimum concentration for each factor in 20μl ISSR-PCR reaction system were 10×Buffer（ Mg2＋ ）2μl, dNTPs 0.4 retool/L, Tag enzyme 1.0 U, primer 0.3μmol/L, DNA30ng.