江西农业大学学报2012，Vol.34Issue(1)：158-164,7.
基于毛细管电泳技术的牡丹SRAP-PCR反应体系优化及引物筛选
Optimization of SRAP-PCR System and Selection of Primers for Paeonia suffruticosa Andr.Based on Capillary Electrophoresis
摘要
Abstract
In this study, the procedure of SRAP - PCR was determined ; an initial denaturing step was performed at 94 X for 5 min; followed by 5 cycles at 94 X for 1 min, 33 X for 1 min and 72 X for 1 min; followed by 35 cycles at 94 X for 1 min, 50 X for 1 min, 72 X for 1 min, and a final extension at 72 X for 10 min. The orthogonal design was used to optimize SRAP-PCR system with five factors at three levels respectively based on capillary electrophoresis. The results showed that the optimized SRAP reaction mixtures for Paeonia suffruticosa Andr. ; (total volume 25 jxL) 50 ng DNA template, 0. 25 mmolL dNTP, 2. 5 mmol/L Mg + , 0.4 (xmol/Lprimer, 0. 5 U Taq DNA polymerase. The concentration of dNTP had the greatest effect and DNA template had the least effect on the result. And twenty-six pairs of primers were selected with abundant polymorphism above 70% from 756 pairs of primers. The optimized SRAP-PCR system and pairs of primers could be applied to related research of Paeonia suffruticosa Andr.关键词
牡丹/SRAP/毛细管电泳/体系优化/引物筛选Key words
Paeonia suffruticosa Andr. / SRAP marker/ capillary electrophoresis/ optimization/ selection of primers分类
农业科技引用本文复制引用
史倩倩,王雁,周琳,黄国伟..基于毛细管电泳技术的牡丹SRAP-PCR反应体系优化及引物筛选[J].江西农业大学学报,2012,34(1):158-164,7.基金项目
国家林业局林业公益性行业科研专项(200904050) （200904050）
