摘要
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℃.
出处
《吉林农业科学》
CSCD
北大核心
2014年第6期21-24,共4页
Journal of Jilin Agricultural Sciences
基金
吉林省重大科技攻关专项(20130204040NY)
吉林省农业创新工程项目(2013~2017)
关键词
花生
SSR
技术体系
分子标记
优化
Peanut
SSR
Reaction system
Molecular markers
Optimization