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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (1): 40-43.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

雷公藤叶片DNA提取及RAPD反应体系建立

林光美1,随粉粉1,林楠2,郑翔宇3,侯长红4   

  • 收稿日期:2009-08-17 修回日期:2009-09-12 出版日期:2010-01-05 发布日期:2010-01-05
  • 基金资助:

    福建省社会发展重点项目;福建省示范推广重点项目

Optimization of RAPD Reaction System on Extraction of Genomic DNA of the leaves of Tripterygium Wilfordii Hook.

  • Received:2009-08-17 Revised:2009-09-12 Online:2010-01-05 Published:2010-01-05

PDF (PC)

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摘要：

摘要］目的：筛选并建立雷公藤叶片基因组DNA的RAPD（Random Amplified Polymorphic DNA）技术最优反应体系。方法：采用改良的CTAB法提取叶片总DNA，然后通过对RAPD反应体系中最主要的退火温度、引物浓度、dNTP浓度、DNA模板浓度以及Tag DNA聚合酶浓度等5个因子逐个进行单因子优化。结果：20μl PCR反应体系中，雷公藤叶片基因组DNA RAPD最优反应体系为，最佳温度36℃，引物浓度为1.1μmol/L，dNTP浓度1.25mmol/L，DNA模板浓度3790 ng/L，Tag DNA聚合酶用量为5.625 U/L。

关键词: 乡村, 乡村, 灾害感知

Abstract:

[Abstract] [Objective] The research aimed to screen out optimum RAPD reaction system on genomic DNA of the leaves of Tripterygium Wilfordii Hook..[Method] The improved CTAB DNA extraction method，to extracted the genomic DNA of the leaves of tripterygium wilfordii Hook. The single factor experiment was adopted to select the required 5 annealing temperature，dNTPs concentration，template DNA concentration，Mg2+ concentration，Taq enzyme concentration and primer concentration.[Result] The optimum reaction system for RAPD amplification of the leaves of Tripterygium Wilfordii Hook. was listed as follows：3790 ng/L DNA，1.1μmol/L primer，1.25mmol/L dNTPs and 5.625 U/L Taq enzyme, the optimum temperature is 56.8℃.