摘要
应用L16(44)正交实验优化番茄EST-SSR标记PCR反应体系。结果表明:20μL体系中各反应物最适浓度为:DNA模板45ng/20μL,引物0.75μmol/L,Mg2+2.0mmol/L,dNTPs0.4mmol/L,Taq DNA酶0.5U/20μL,且Mg2+对该标记反应体系影响最大。利用5对EST-SSR引物及P1、P2基因组DNA验证优化体系的可靠性,为该标记在番茄基因组分子标记辅助育种提供了试验依据。
L16（44） orthogonal design was used in the EST-SSR Maker PCR system optimization of tomato.The results showed that the best PCR system was：45 ng/20μL DNA template,0.75 μmol/L primer,2.0 mmol/L Mg2＋,0.4 mmol/L dNTPs,0.5 U/20μL Taq DNA polymerase,and the effect of the primer concentration was the greatest for this system.The system had been verified by five pairs of primers and P1,P2 genome DNA,and provided experiments reference on molecular marker assisted selection of tomato.
出处
《北方园艺》
CAS
北大核心
2011年第16期142-144,共3页
Northern Horticulture
基金
国家“863”计划资助项目(2006AA100108-3-1)
山东省农业良种工程资助项目(2010LZ006-01)
