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中国农学通报 ›› 2013, Vol. 29 ›› Issue (36): 319-325.doi: 10.11924/j.issn.1000-6850.2013-0490

所属专题: 生物技术 油料作物

• 生物技术科学 • 上一篇    下一篇

大豆不同生长时期基因组DNA提取方法的优化

王月月 范龙 张洁 王冬梅   

  • 收稿日期:2013-02-26 修回日期:2013-04-18 出版日期:2013-12-25 发布日期:2013-12-25
  • 基金资助:
    转基因生物新品种培育重大专项子课题;河北省教育厅项目

Optimization of DNA extraction methods from soybean in different period

  • Received:2013-02-26 Revised:2013-04-18 Online:2013-12-25 Published:2013-12-25

摘要: 以优质大豆品种冀豆7号、冀豆16号、五星1号、五星2号和MK(Maverick)不同生长时期的叶片为材料,分别采用不同方法提取大豆基因组,并通过对提取的DNA的含量测定、琼脂糖凝胶电泳分析和内参基因的PCR扩增,对不同方法所提取的DNA质量进行综合评价。结果表明:在嫩叶期,改进的CTAB法能够 分离出纯度高、质量好的DNA,而在开花期,采用本试验的新改良CTAB法可以有效去除多糖、酚类等物质得到完整性和纯度较好的大豆基因组,为大豆的分子生物学研究奠定了基础。

关键词: 解剖, 解剖

Abstract: Using leaves in different growth periods of high-quality soybean varieties J-17, J-16, WX-1, WX-2 and Maverick as experimental materials, we used different methods for the genomic DNA extraction and determined the concentration of genome DNA. The DNA were tested by agarose gel electrophoresis, PCR amplification for internal control and comprehensive evaluation of the quality of different methods for DNA extraction. The results indicated that the DNA extracted with the modified CTAB method had higher quality and purity in fresh leaves period. The most of polysaccharides and polyphenolic compounds could be removed efficiently by utilizing new modified CTAB method and the new method was suitable for extracting high qualified DNA in flowering stage. This study laid the foundation of soybean moleculer bioresearch.