水稻SSR-PCR的反应体系优化及RIL亲本间多态性引物筛选

Optimization of Rice SSR-PCR Reaction System and Screening of Polymorphism Primer with Parents Characteristics

为比较三组概率分析法和传统QTL定位法,优化了水稻Ⅱ-32B和特青构建的F7重组自交系(Recombinant inbred line,RIL)后代群体SSR-PCR的反应体系,筛选出在双亲间具有多态性的引物。结果表明:适宜水稻的SSR-PCR反应体系(10μL)为10×Buffer 2μL,Mg2+终浓度2.0mmol/L,dNTP终浓度0.5mmol/L,引物终浓度1.0μmol/L,DNA模板1μL,Taq DNA聚合酶0.15U。从分布于全基因组的SSR引物中筛选出142对在双亲间具有多态性的引物,为完成后续的QTL定位研究奠定良好的基础。

The rice SSR-PCR reaction system of F7population of recombinant inbred line from U-32BX Teqing was optimized by Tri-group Probability analysis and traditional QTC analysis to screen polymorphismic primers with parents characteristics. The results showed that the suitable rice SSR PCR reaction system included 2μL 10×Buffer, 2.0 mmol/L Mg2＋ , 0.5 mmol/L dNTP, 1.0 μmol/L primer, 1μL DNA template and 1μL Taq DNA polymerase. 142 pairs of polymorphismic primers with parents characteristics were screened from SSR primers distributed in the total genome, which lay a good fundation for subsequent OTL analysis.