花生基因组SRAP-PCR体系的优化

Optimization of SRAP-PCR Reaction System in Peanut Genome

【研究目的】研究花生基因组SRAP-PCR的扩增条件,建立其优化扩增体系;【方法】对模板DNA、引物、dNTP mixture、Taq DNA聚合酶浓度及退火温度设置不同梯度,研究各因素对PCR结果的影响;【结果】扩增程序为:94℃变性2min,1Cycle;94℃变性30s,48℃复性30s,72℃延伸1min,40Cycles;72℃延伸7min。反应体系成份为:DNA模板80ng,左右引物各200ng,dNTP mixture2.0mmol/L,Taq DNA聚合酶3U,10×Buffer8μl,总体积60μl。【结论】建立了满足花生基因组SRAP-PCR的优化扩增体系。

[OBJECTIVE] Study optimum condition for SRAP-PCR in peanut genome and set up optimum system. [METHOD] Different levels on concentration of DNA template, primer, dNTP mixture and Taq DNA polymerase and annealing temperature were set in this experimentation. All factor influence for SRAP-PCR in peanut genome was investigated. [RESULTS] A optimum system that could satisfy SRAPPCR in peanut was established. The amplification program was： 94℃ for 30s, 48℃ for 30s, 72℃ for lmin, 40cycles, 72℃ for 7min. The 60μl reaction mixture contained： DNA template 80ng, each primer 200ng, dNTP mixture 2.0mmol/L, Taq DNA polymerase 3U, 10× Buffer 8μl. [CONCLUSION] Optimum system for SRAP-PCR in peanut genome was set up.