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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (18): 57-60.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

太行花DNA提取的优化和适用分子标记检测

方向民,王红卫,程月琴,叶永忠,杨 程   

  • 收稿日期:2009-04-16 修回日期:2009-05-11 出版日期:2009-09-20 发布日期:2009-09-20
  • 基金资助:

    河南农业大学博士科研启动基金

Optimization of total DNA extraction and test of suitable molecular markers in Taihanggia rupestris

  • Received:2009-04-16 Revised:2009-05-11 Online:2009-09-20 Published:2009-09-20

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摘要：

比较了常规CTAB法、改良CTAB法和SDS法对太行花叶片总DNA的提取效果，并对改良CTAB法提取的DNA在多种分子标记中的适用性进行了测试。结果表明：常规CTAB法提取的DNA难以完全溶解，且有褐化现象；SDS法提取的DNA产率及纯度都很低；改良CTAB法提取的DNA产率高且稳定，无明显降解，杂质少，OD260/OD280比值在1.8左右。以改良CTAB法提取的DNA为模板，应用叶绿体和线粒体通用引物扩增出了特异性的高效产物，ISSR和RAPD引物对总DNA的扩增也获得理想结果。因此，改良CTAB法适用于太行花总DNA提取，其产物能满足核、叶绿体和线粒体基因组分子实验的要求。

关键词: 烟粉虱成虫, 烟粉虱成虫, 空间格局, 时序动态, 黄瓜

Abstract:

The total DNA was isolated from the leaf of Taihanggia rupestris YU et LI by the conventional CTAB method，improved CTAB method and SDS method. The extracts by improved CTAB method were tested as the template for several molecular marker types．The results showed the extracts by conventional CTAB method were brown and could not dissolve completely, and those by SDS method had a low quality and yield. As for the improved CTAB method, there was a high production rate of the extracts，that was pure with little degradation，and A260/ A280 was 1.8 or so. With these leaf DNA serving as template, PCR at two loci of mitochondrial and chloroplast genomes was effective and special highly. Besides, these total DNA were good templates for SSR and RAPD molecular markers. So improved CTAB method is a good protocol by which the total DNA was isolated from the leaf of T. rupestris, its product meet the requirement of PCR amplification for the nuclear, chloroplast and mitochondrial genomes.