四川农业大学学报 ›› 2010, Vol. 28 ›› Issue (04): 467-470.doi: 10.3969/j.issn.1000-2650.2010.04.014

• 研究论文 • 上一篇    下一篇

披碱草属物种AFLP体系优化

陈仕勇, 马啸, 齐晓芳, 张新全   

  1. 四川农业大学动物科技学院, 四川 雅安 625014
  • 收稿日期:2009-07-25 出版日期:2010-12-31 发布日期:2017-03-01
  • 通讯作者: 张新全(E-mail:zhangxq@sicau.edu.cn) E-mail:zhangxq@sicau.edu.cn
  • 基金资助:
    国家重点基础研究973计划(2007CB108907);国家自然科学基金项目(31072077)

Optimization of AFLP in Elymus (Poaceae)

CHEN Shi-yong, MA Xiao, QI Xiao-fang, ZHANG Xin-quan   

  1. College of Animal Science and Technology, Sichuan Agricultural University, Yaan 625014, Sichuan, China
  • Received:2009-07-25 Online:2010-12-31 Published:2017-03-01

摘要: 试验对内切酶EcoRⅠ和MseⅠ的用量和酶切时间、预扩和选扩过程中模板稀释倍数以及扩增引物浓度分别进行了优化,并采用了Bassam法和Sanguinetti银染法进行了比较分析。结果表明:采用3UEcoRⅠ和MseⅠ这两种酶在37℃下酶切3h,预扩模板稀释10倍和1.5ng/μL引物进行预扩,选扩模板稀释20倍和2ng/μL引物进行选扩,采用Bassam法进行银染,能够得到较好的试验结果,为AFLP进一步在披碱草属中的应用提供了参考。

关键词: 披碱草属, 扩增片段长度多态性标记(AFLP), 银染, 体系优化

Abstract: The key factors affecting the AFLP analysis in Elymus were optimized,including the digesting time and the content of the EcoRⅠ and MseⅠ,diluted ratio and concentrations of primers in the pre-amplification and selective amplification.Furthermore,the methods of Bassam and Sanguinetti silver staining were also compared in the staining quality.The optimal reaction system of AFLP were established,which including digest with 3 units of each enzymes at 37℃ for 3 h,pre-amplification performed with templates diluted 10 times and 1.5 ng/μL primers,selective amplification performed with templates diluted 20 times and 2 ng/μL primers,as well as stained with Bassam silver staining.The results could apply for the further study on AFLP in Elymus.

Key words: Elymus, AFLP, silver staining, optimization

中图分类号: 

  • S543.024