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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (9): 38-41.

所属专题： 生物技术； 油料作物

• 农学 农业基础科学 • 上一篇    下一篇

一种改良的快速高质大豆基因组DNA提取方法

任良真 张春宝 赵洪锟 董英山 赵丽梅   

  • 收稿日期:2011-10-08 修回日期:2011-12-06 出版日期:2012-03-25 发布日期:2012-03-25
  • 基金资助:

    国家科技部“863”计划重点项目“强优势大豆杂交种的创制与应用”;国家农业部转基因生物新品种培育科技重大专项“抗病虫转基因大豆新品种培育”

An Improved Method to Rapid and High-quality of Genomic DNA Extraction from Soybean

  • Received:2011-10-08 Revised:2011-12-06 Online:2012-03-25 Published:2012-03-25

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摘要：

为了探索一种适用于大豆叶片基因组DNA的快速、高效的提取方法，在传统的SDS提取方法的基础上，通过将缓冲液成分和浓度进行改良并加入表面活性剂Tween-20，优化实验流程。用限制性内切酶对该方法提取的DNA进行酶切及电泳检测，可得均一弥散条带。以提取的DNA模版扩增大豆Actin基因，测序后证实片段正确。同样以其为模版以SSR引物Sctt011进行SSR-PCR反应，并通过聚丙烯酰胺凝胶电泳检测，也可观察到特异性明显且无杂质的目的条带。实验结果表明，该方法提取的DNA完全符合各种分子实验的要求。本研究方法可用于快速提取大豆叶片基因组DNA，同时提高了提取DNA的质量。

关键词: 安徽省, 安徽省

Abstract:

In order to explore a method to fast extract high-quality soybean leaf genomic DNA. This study improved buffer composition and concentration, added surfactant Tween-20, and optimize the experimental procedure on the basis of the SDS method. The extracted DNA was digested with the restriction enzyme EcoR I, electrophoresis displayed it was digested completely. And the DNA was used as template to amplify the soybean Actin gene, sequencing confirmed the segment was correct. And the DNA was also used for SSR-PCR reaction with SSR primer Sctt011, polyacrylamide gel electrophoresis results showed the bands were specifically and clearly. The results show that the extracted DNA can be used for various molecular experiments. The method can shorten the DNA extraction time, while improve the quality of extracted DNA.

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