›› 2009, Vol. 17 ›› Issue (3): 354-357.DOI: 10.11733/j.issn.1007-0435.2009.03.017

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Establishment and Optimization of ISSR Reaction System for Agropyron cristatum(L.) Gaertn.

WANG Fang1, YUAN Qing-hua1,2   

  1. 1. College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou, Gansu Province 730020, China;
    2. Insfitute of Animal Science, CAAS, Beijing 100193, china
  • Received:2008-11-09 Revised:2009-03-11 Online:2009-06-15 Published:2009-06-15

冰草ISSR-PCR反应体系的建立与优化

王方1, 袁庆华1,2   

  1. 1. 兰州大学草地农业科技学院, 兰州, 730020;
    2. 中国农业科学院北京畜牧兽医研究所, 北京, 100193
  • 通讯作者: 袁庆华,E-mail:yuanqinghua@hotmail.com
  • 作者简介:王方(1983- ),女,汉族,甘肃兰州人,硕士研究生,研究方向为牧草种质资源,E-mail:wfcayenne@163.com
  • 基金资助:
    国家科技支撑项目(2008BABD3B01);基本科研业务费专项(YWF-TD-3)

Abstract: It is necessary to establish and optimize the ISSR-PCR reaction system for genetic diversity study of Agropyron cristatum(L.) Gaertn.Results show that the optimum concentration of seven reactants in 20 μL reaction mixture were as follows: 40 ng genomic DNA,0.2 mmol·L-1 dNTP,0.5 μmol·L-1 ISSR primer,1 U Taq DNA polymerase,2 μL 10× PCR buffer,2.5 mmol·L-1 MgCl2.The suitable annealing temperature of primer 815 was 50℃.This reaction system would provide the basis for the diversity analysis of A.cristatum with ISSR markers.

Key words: Wheatgrass, ISSR, PCR reaction system, Optimization

摘要: 为进一步开展冰草属植物种质资源遗传多样性研究,寻找可用于冰草(Agropyron cristatum(L.)Gaertn.)的ISSR-PCR最适反应体系并建立与优化,具有重要的意义。通过优化影响ISSR-PCR体系的主要参数,最终确定在20μL体系中各反应物的最适含量为:40 ng模板DNA,0.2 mmol·L-1dNTP,0.5μmol·L-1ISSR引物,1 UTaq DNA聚合酶,2μL10×PCR Buffer,2.5 mmol·L-1MgCl2;引物815号的最适退火温度为50℃。该体系的建立为今后利用ISSR技术进行冰草属种质资源的遗传多样性分析奠定了技术基础。

关键词: 冰草, ISSR, PCR反应体系, 优化

CLC Number: