蚕豆SSR-PCR体系建立及优化

Establishment and optimization of SSR-PCR system in faba bean

为了建立适宜蚕豆的SSR-PCR反应体系,用于蚕豆的SSR分子标记研究。以青海12号、透心绿为材料,通过正交设计L16(45)对蚕豆SSR-PCR反应体系中的Taq酶,Mg^(2+),dNTPs,引物和模板DNA 5个因素的4个水平进行优化试验。结果表明:各因素不同水平对PCR反应均有影响,各因素对PCR扩增结果的影响大小分别为:Taq酶>dNTPs>模板DNA>引物>Mg2+。最终筛选出蚕豆最佳SSR-PCR反应体系(20μL):1U Taq酶,0.75 mmol/L d NTPs,100 ng模板DNA,2μmol/L引物和1.5 mmol/L Mg^(2+)。

The variety of Qinghai No.12 and Touxinlv were chosen as experimental materials in order to establish the appropriate SSR-PCR system for analyzing SSR molecular marker of faba bean. The SSR-PCR system of faba bean were optimized by using the orthogonal design L_(16)( 4~5) for Taq DNA polymerase,Mg^(2+),dNTPs,primer and DNA template. The results showed that different levels of concentration of various were significantly affected the results of PCR. And the effect size is Taq DNA polymerase > d NTPs > DNA template > primer > Mg^(2+). Finally,an optimal 20μL volumes reaction system of faba bean consisted of 1U Taq DNA polymerase,0. 75 mmol/L d NTPs,100 ng DNA template,2μmol/L primer and 1. 5 mmol/L Mg^(2+).