安徽农业大学学报2013，Vol.40Issue(3)：387-392,6.
杜仲SSR-PCR反应体系建立及引物筛选
Establishment of SSR-PCR reaction system and primers screening for Eucommia ulmoides
摘要
Abstract
To provide theoretic and technical support for molecular assessment of genetic diversity in germplasm resources of Eucommia ulmoides,simple sequence repeat-polymerase chain reaction (SSR-PCR) system was established through orthogonal test and optimized through single factor experiment.The results showed that the optimal SSR-PCR reaction system was a volume of 10.0 μL containing forward-and reversed-primer (10 μmol·L-1) 0.2 μL,respectively,Taq DNA polymerase (5 U·μL-1) 0.1 μL,dNTP (10 mmol·L-1) 0.2 μL,DNA (5-10ng·μL-1) 1.0 μL,Mg2+ (25 mmol·L-1) 0.6 pL,10 × PCR buffer 1.0 pL and HPLC water 6.7 μL.Three SSR polymorphic primer pairs were selected.The results proved the limited transferability of SSR primers between distantly related species,revealed the genetic diversity of E.ulmoides preliminarily,and exhibited the advantages of SSRs for the investigation of genetic diversity.关键词
杜仲/微卫星标记/PCR反应体系/引物筛选/遗传多样性Key words
Eucommia ulmoides/ SSRs/ PCR reaction system/ primer screening/ genetic diversity分类
农业科技引用本文复制引用
朱晓敏,王弦云,陈龙灿,康向阳,王勤,束庆龙,张良富,曹翠萍..杜仲SSR-PCR反应体系建立及引物筛选[J].安徽农业大学学报,2013,40(3):387-392,6.基金项目
安徽高校省级自然科学重点项目 (KJ2011A115)资助. （KJ2011A115）
