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Establishment and Optimization of SSR-PCR Reaction System in Potato

ZHANG Feng-jun1,2,ZHANG Yong-cheng1,2 (1.Qinghai Academy of Agriculture and Forestry Science,Xining Qinghai 810016,China;2.The Key Laboratory for Potato Breeding of Qinghai Province,Xining Qinghai 810016,China)  
Used L16(45) 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,Mg2+ 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 ℃.
【Fund】: 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)
【CateGory Index】: S532
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