花生SSR分子标记技术体系的建立和优化

Establishment and Optimization of SSR Molecular Marker Technology System in Peanut

本试验以8份花生种质资源为试材,摸索适宜的DNA提取方法,并通过大量文献的调研初步筛选出4套反应体系,并使用不同引物对其进行筛选,最终确立了适合花生SSR-PCR反应体系为10μL,其中各组分终浓度如下:25 ng DNA,1×buffer,0.15μmol/L引物,200μmol/L d NTP,0.5UTaq酶。最终确立的扩增程序为:94℃预变性5 min,94℃变性1 min,55℃退火30 s,72℃延伸1 min,35个循环,72℃延伸5 min。

Using eight species of peanut germplasm resources as the experimental materials in this experiment, the appropriate DNA extraction method was explored. 4 sets of reaction system were selected through reading a large amount of papers and preliminary screening. Different primers were screened in the reaction system. SSR-PCR system suitable to peanut was finally established, in which for 10 μL, 25ng DNA, lbuffer, 0.15μmol/L primer, 200μmol/ L dNTP, and 0.5UTaq. A PCR system was finally established, in which pre-denaturing 5 min at 94 ℃, denaturing for 1 min at 94℃, annealing for 30 s at 55℃, extending for 1 min at 72℃, 35 cycles, extending for 5 min at 72℃.