四川农业大学学报 ›› 2006, Vol. 24 ›› Issue (03): 360-363.doi: 10.16036/j.issn.1000-2650.2006.03.024

• 研究简报 • 上一篇    下一篇

珍稀濒危竹种筇竹(Qiongzhuea tumidinoda)RAPD反应条件的优化

王波, 高素萍, 陈其兵   

  1. 四川农业大学 林学园艺学院, 四川 雅安 625014
  • 收稿日期:2006-03-22 出版日期:2006-09-30 发布日期:2017-03-06
  • 基金资助:
    四川省教育厅项目(2004A030)

Optimization of RAPD Conditions of a Rare and Endangered Bamboo of Qiongzhuea tumidinoda

WANG Bo, GAO Su-ping, CHEN Qi-bing   

  1. College of Forestry and Horticulture, Sichuan Agricultural University, Yaan 625014, Sichuan, China
  • Received:2006-03-22 Online:2006-09-30 Published:2017-03-06

摘要: 以宜宾筇竹为材料建立了筇竹RAPD反应优化体系,用于筇竹遗传多样性分析,以改进的CTAB法提取筇竹叶片总DNA,分别测试了镁离子浓度、dNTP浓度、引物浓度、模板DNA含量、DNA聚合酶量对反应结果的影响。通过对各因子的组合比较,建立了筇竹RAPD反应优化体系:20μL PCR反应体积,10×Taq酶配套缓冲液(2μL);1.25U Taq酶;75 ng模板DNA;45 ng引物;1.88 mmol/L MgCl2;0.19 mmol/L dNTP。

关键词: 筇竹, 遗传多样性, 优化, CTAB, RAPD

Abstract: The Qiongzhuea tumidinoda RAPD reaction system optimized with yibin is established in ordered to be used in Qiongzhuea tumidinoda hereditary variety analysis. The genomic DNA of Qiongzhuea tumidinoda is extracted with improved CTAB method. The effect of content of Mg2+, dNTP, DNA templates, primers and DNA polymerase on experimental results is tested and the optimal reaction system of RAPD Qiongzhuea tumidinoda is determined as the following: 10×Taq polymerase corresponding buffer(2 μL), 1.25 U Taq polymerase, 75 ng template DNA, 45 ng primer, 1.88 mmol/L MgCl2, 0.19 mmol/L dNTP in total 20 μL reaction volume. This is to use RAPD marker to settle solid foundation for the classification of Qiongzhuea tumidinoda again on the molecular level.

Key words: Qiongzhuea tumidinoda, hereditary variety analysis, optimization, CTAB, RAPD

中图分类号: 

  • S795