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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (14): 90-93.

• 生物技术科学 • 上一篇    下一篇

淮山RAPD-PCR反应体系的建立

周鑫 程文杰 罗 欣 黄小龙 陈吉良 黄祖旬 吴文嫱 黄东益   

  • 收稿日期:2010-03-26 修回日期:2010-04-15 出版日期:2010-07-20 发布日期:2010-07-20

The establishment of RAPD-PCR system of Dioscorea opposite Thunb

  • Received:2010-03-26 Revised:2010-04-15 Online:2010-07-20 Published:2010-07-20

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摘要： 摘要：采用改良CTAB法提取淮山的叶片总DNA。通过单因子实验法分析DNA浓度、引物浓度、dNTP浓度、Taq聚合酶以及Mg2+浓度对RAPD-PCR扩增结果的影响，建立了适合于淮山RAPD-PCR反应体系，即20μL体系中包括：模板DNA为50 ng、Taq酶为0.2U、Mg2+浓度为3.0mmol/L、引物浓度为0.2μmol/L、dNTPs浓度为0.2mmol/L。

关键词: 生态环境建设, 生态环境建设, 生态学理论, 城市园林绿化, 恢复生态学

Abstract: abstract: The total DNAs of Dioscorea opposite Thunb were extract by the improved CTAB method, and set the gradient by single factor experiment with template DNA concentration, Primer concentration, dNTP concentration, Taq concentration and Mg2+ concentration, Study on the effect of single-factor change on D. opposite RAPD-PCR amplification. In conclusion, the D. opposite reaction system was optimized as below: 50ng template DNA, 0.3U Taq DNA polymerse, 3.0mmol/L Mg2+,0.2μmol/L primer and 0.2mmol/L dNTPs with total 20μL reaction solution.