安徽农业科学2012，Vol.40Issue(28)：13709-13712,13719,5.
老芒麦ISSR-PCR反应体系的建立与优化
Establishment and Optimization of ISSR Reaction System for Elymus sibiricus L.
摘要
Abstract
[Objective] This study aimed to establish and optimize the ISSR-PCR reaction system and amplification process for Elymus sibirkus L. , to provide scientific basis for exploring the genetic diversity of E. sibiricus germplasm resources. [Method] Orthogonal design and single factor test were applied to establish the ISSR-PCR reaction system of E. sibiricus, optimize the affecting factors including Taq DNA polymer-ase, template DNA concentration, Mg2+ , dNTP, primer concentration, and screen the annealing temperature, number of cycles and extension time. [ Result] The optimal reaction system for ISSR analysis contains 0. 2 mmol/L dNTPs ,0.2 μmol/L ISSR primers, 1. 5 U of Taq DNA polymerase, 2.5 μl of 10 X PCR Buffer ,1.5 mmol/L MgCl2 and 40 ng of template DNA in 25 Μl total volume; the amplification was conducted with 35 cycles and extension time of 90 s. [Conclusion] ISSR-PCR reaction system for E. sibiricus L. was established and optimized in this study, which was verified using two E. sibiricus germplasms, results show that the ISSR-PCR reaction system is stable and can be used for the genetic analysis of E. sibirkus.关键词
老芒麦/ISSR/正交试验/单因素试验Key words
Elymus sibirkus L/ ISSR/ Orthogonal experiment/ Single factor test分类
农业科技引用本文复制引用
周国栋,李志勇,扈顺,李鸿雁,师文贵,刘磊..老芒麦ISSR-PCR反应体系的建立与优化[J].安徽农业科学,2012,40(28):13709-13712,13719,5.基金项目
多年生牧草种质资源收集、编目与利用项目(NB2012-2130-135-33) （NB2012-2130-135-33）
牧草种质资源保护项目. （）
