植物学报 ›› 2002, Vol. 19 ›› Issue (04): 452-456.

• 研究论文 • 上一篇    下一篇

濒危植物七子花RAPD条件的优化

李钧敏 金则新 柯世省   

  1. (台州师范专科学校生化系 浙江临海 317000)
  • 收稿日期:2001-07-06 修回日期:2001-08-03 出版日期:2002-07-20 发布日期:2002-07-20
  • 通讯作者: 李钧敏

Optimal Choice for Compositions in RAPD Analysisof Heptacodium miconioides

LI Jun-Min JIN Ze-Xin KE Shi-Sheng   

  1. (Biology and Chemistry Department, Taizhou Teachers' College, Linhai, Zhengjiang 317000)
  • Received:2001-07-06 Revised:2001-08-03 Online:2002-07-20 Published:2002-07-20
  • Contact: LI Jun-Min

摘要: 以改进SDS法抽提濒危植物七子花嫩叶总DNA,进行随机扩增多态DNA(RAPD)分析,分别测试了镁离子, dNTP,模板DNA含量,引物和DNA聚合酶量对反应结果的影响,通过各因子的组合研究,可知七子花RAPD分析较适宜的扩增条件是:15 μL PCR反应体积,1×Taq酶配套缓冲液(10 mmol/L Tris·HCl pH 9.0, 50 mmol/L KCl, 0.1% Triton X_100),2.5 mmol/L MgCl2,2U Taq酶(上海华美公司),10 ng模板DNA,20 pmol引物(上海Sangon公司);dATP、dCTP 、dGTP 、dTTP 各0.1 mmol/L。

Abstract: The DNA templates of Heptacodium miconioides extracted from young leaf were amplified by RAPD. Effects of the content of Mg2+, dNTP, DNA templates, primers and DNA polymerase on experimental results were tested and the optimal reaction system of RAPD for Heptacodium miconioides was determined as follows: 1×Taq polymerase corresponding buffer (10 mmol/L Tris·HCl pH 9.0,50 mmol/L KCl, 0.1% Triton X_100), 2.5 mmol/L MgCl2, 2U Taq DNA polymerase, 10 ng template DNA, 20 pmol primer, 0.1 mmol/L dATP, dCTP, dGTP, dTTP separately in total 15 μL reaction volume.