糯玉米SSR-PCR反应体系优化及多态性引物筛选  

Optimization of SSR- PCR System of Waxy Maize and Selection of Polymorphic Primers

作　　者：王薪淇[1] 姜龙[1] 宋轶群[1] 邢政[1] 储航[1] 赵仁贵[1] 

机构地区：[1]吉林农业大学农学院,长春130118

出　　处：《吉林农业大学学报》2016年第4期398-404,共7页Journal of Jilin Agricultural University

基　　金：国家星火计划项目(2011GA660005);吉林省财政厅科研育种项目(吉财2012002);长春市科技支撑计划项目(长科技合2011215号)

摘　　要：用L25(56)正交设计建立和优化糯玉米SSR-PCR反应体系;利用370对SSR引物对糯玉米两亲本及F1进行扩增,筛选抗丝黑穗病的相关引物。结果表明:20μL糯玉米SSR-PCR最优反应体系为1.5 U DNA聚合酶,1.0 nmol/μL镁离子,80 ng模板DNA,0.1 pmol/μL引物,0.15μmol/μL d NTPs,1倍扩增缓冲液;370对SSR引物对两亲本及F1进行扩增的条带显示,有差异且条带清晰的引物为87对。确立了糯玉米SSR-PCR最优反应体系,筛选并得到了87对多态性引物。SSR-PCR system of waxy maize was established and optimized with L25（56） orthogonal design. Parents and F1 of waxy maize were amplified with 370 pairs of SSR primers and primers resistant to head smut were selected. The results showed that the optimal SSR-PCR system of waxy maize in a total volume of 20 μL is template 1.5 U DNA polymerase, 1.0 nmol/ixL Mg2＋, 80 ng DNA template, 0. 1 pmol/μL primers, 0. 15 μmol/μL dNTPs, and 1xPCR buffer; The parents and FI were amplified with 370 pairs of SSR primers, and 87 pairs were selected based on their discrepant and clear banding patterns.

关 键 词：糯玉米 SSR—PCR 正交设计 引物筛选 

分 类 号：S330.2[农业科学—作物遗传育种]