华北农学报2016，Vol.31Issue(4)：68-73,6.DOI:10.7668/hbnxb.2016.04.012
绣球 SSR-PCR 反应体系的建立与优化
Establishment and Optimization of SSR-PCR Reaction System in Hydrangea macrophylla
摘要
Abstract
In order to establish and optimize the SSR-PCR reaction system of Hydrangea macrophylla,and the orthogonal design L25 (5 6 )was applied to optimize the five main factors(Mg2 +,dNTPs,primers,DNA template,and Taq enzyme)at five levels of SSR-PCR reaction system.The PCR result was analyzed to select the best levels of five factors and established the suitable SSR reaction system for Hydrangea macrophylla.The results showed that the op-timal SSR-PCR system of 20 μL volumes consists of 60 ng DNA template,1 .5 mmol /L Mg2 +,0.3 mmol /L dNTPs, 0.4 μmol /L primer,and 0.8 U Taq polymerase.PCR amplification procedures were pre-denaturation for 5 min at 94 ℃;followed by 33 cycles of denaturation for 1 min at 94 ℃,annealing at a suitable for primers for 40 s,exten-sion for 1 min at 72 ℃;and a final extension for 10 min at 72 ℃,then stored at 4 ℃.The optimal SSR-PCR reac-tion system was used to verify by 10 cultivars of Hydrangea macrophylla,and the results showed that it was excellent stability and repeatability for the optimal reaction system.The establishment of this reaction system provides a theo-retical basis and technical references for further research on the genus Hydrangea by SSR markers.关键词
绣球/SSR-PCR/正交设计/反应体系Key words
Hydrangea macrophylla/SSR-PCR/Orthogonal design/Reaction system分类
农业科技引用本文复制引用
贾新平,孙晓波,梁丽建,邓衍明,苏家乐,周建涛..绣球 SSR-PCR 反应体系的建立与优化[J].华北农学报,2016,31(4):68-73,6.基金项目
江苏省科技支撑计划项目（BE2014408）；江苏省农科院成果转化与推广项目（KF15（5005）） （）
