Optimization of SSR-PCR Reaction System and Screening of Polymorphic Primers for RIL Parents

水稻SSR-PCR的反应体系优化及RIL亲本间多态性引物筛选(英文)

[Objective] This study aimed to establish a high-efficiency and stable SSR amplification system and screen polymorphic primers in order to further compare the tri-group probability analysis method and traditional QTL mapping method. [Method] In this study, we preliminarily screened the 605 pairs of primers evenly distributed on the 12 chromosomes through investigating their polymorphism performance in amplification, and established an optimized SSR-PCR reaction system. [Result] A 10μL SSR-PCR reaction system suitable for rice was set up as fol ows： 2 μl of 10 × Buffer, 2.0 mmol/L Mg2＋ （final concentration）, 0.5 mmol/L dNTPs, 1.0 μmol/L primers （final concentration）, 1 μl of DNA template, 0.15 U Taq DNA polymerase. Among the SSR primers distributed over the genome, 142 pairs that were polymorphic upon the parents were screened. [Conclusion] This study lays a good foundation for sub-sequent QTL mapping studies.

[目的]为比较三组概率分析法和传统QTL定位法,建立高效稳定的SSR扩增技术体系,筛选出在双亲间具有多态性的引物。[方法]本研究利用水稻Ⅱ-32B和特青两个亲本对605对均匀分布于12条染色体的SSR引物的扩增情况进行了初步筛选,并研究了其扩增多态性和最优SSR-PCR反应体系[结果]结果表明:适宜水稻的SSR-PCR反应体系(10μl)为10×Buffer 2μl,Mg2+终浓度2.0 mmol/L,dNTP终浓度0.5 mmol/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。[结论]为完成后续的QTL定位研究奠定良好的基础。