马铃薯SSR-PCR反应体系的建立和优化  

Establishment and Optimization of SSR-PCR Reaction System in Potato

作　　者：张凤军[1,2] 张永成[1,2] 

机构地区：[1]青海省农林科学院,青海西宁810016 [2]青海省马铃薯育种重点实验室,青海西宁810016

出　　处：《种子》2011年第1期8-10,共3页Seed

基　　金：青海省"十一五"重大科技攻关项目:马铃薯新品种选育(2006-N-129);国家科技支撑计划项目:高产优质专用马铃薯育种技术研究及新品种选育(2006BAD01A06-1-10);国家马铃薯产业技术体系西宁试验站资助基金(zhsyz-27)

摘　　要：应用L16(45)正交设计对影响马铃薯SSR-PCR的主要参数进行优化,建立适于马铃薯SSR-PCR反应的最佳体系。结果表明:各因素不同水平浓度对PCR反应结果均有显著影响。马铃薯SSR-PCR优化反应体系为:10×PCR Buffer 1.0μl、模板DNA约30 ng、dNTP 200 mol/L、SSR引物66 ng、Taq DNA聚合酶0.25 U、Mg2+2.25 mmol/L,加ddH2O至终体积10.0μl。PCR扩增程序如下:94℃预变性5 min;94℃变性1 min,53℃复性1 min,72℃延伸1.5 min,共35个循环;72℃延伸10 min,然后4℃保存。Used L16（4^5） orthogonal designs for optimizing the main factors of the SSR-PCR reaction system of potato and establishing the optimum SSR reaction system finally.The result showed that there were significant effects on the results of SSR-PCR at different concentration of each factor,and the optimized SSR-PCR reaction system was：10×PCR Buffer 1.0μl,DNA template 30 ng,dNTP 200 μmol/L,SSR primer 66 ng,Taq DNA polymerase 0.25 U,Mg^2＋ 2.25 mmol/L.,adding ddH2O to terminal volume 10.0 μl.The PCR amplification procedure was：predenaturation for 5 min at 94 ℃,followed by 35 cycles of denaturation for 1 min at 94 ℃,anneal for 1 min at 53 ℃,extension for 1.5 min at 72 ℃,the amplification was completed after extension for 10 min at 72 ℃,then stored at 4 ℃.

关 键 词：马铃薯 正交设计 SSR-PCR 体系优化 

分 类 号：S532[农业科学—作物学]