短芒披碱草SRAP-PCR 体系的建立和优化

通过单因子和正交设计两种试验方法,对短芒披碱草SRAP-PCR体系进行优化,得到短芒披碱草20μL SRAP-PCR最优反应体系为：Mg2＋2.00 mmol/L、引物0.40μmol/L、Taq DNA聚合酶1.0 U、d NTPs250μmol/L、DNA 30 ng和10×buffer 2μL;各因素水平变化对PCR反应影响从大到小依次是：Mg2＋、引物、Taq酶、d NTPs和模板DNA。运用该优化体系对6份短芒披碱草材料进行验证,电泳结果显示扩增条带多态性高,清晰无杂带。该优化体系的建立有助于将SRAP标记技术用于短芒披碱草的遗传育种等研究。

Establishment and Optimization of SRAP-PCR Reaction System for Elymus breviaristatus

The SRAP-PCR for Elymus breviaristatus was optimized using the single factor test and the orthogonal design test in the study. The optimized SRAP- PCR system for E. breviaristatus was： 2.00 mmol/L MgC12, 0.40 μmol/L primers, 1.0 U Taq DNA polymerase, 250 μmol/L dNTPs, 30 ng DNA template, 2 μL 10 × PCR buffer in the total volume of 20 μL. The results showed that the order for the most influenced factor to the least influenced factor was： Mg2＋, primer, Taq polymerase, dNTPs and DNA template. The optimized PCR system was confirmed using six E. breviaristatus accessions. The electrophoresis results showed that the amplified bands were highly polymorphic and clear, further indicating that the optimized system was reliable. The optimized SRAP- PCR system could be used for genetics and breeding studies in E. breviaristatus or other Elvmus species.