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中国农学通报 ›› 2013, Vol. 29 ›› Issue (33): 319-322.doi: 10.11924/j.issn.1000-6850.2013-1064

所属专题: 生物技术

• 生物技术科学 • 上一篇    下一篇

梭梭属2种植物线粒体基因组的提取

吴明朝 张明慧 杨丽丽 石磊 陈虞超 宋玉霞   

  • 收稿日期:2013-04-15 修回日期:2013-05-24 出版日期:2013-11-25 发布日期:2013-11-25
  • 基金资助:
    国家自然科学基金项目

The Study on Extracting Mitochondrial DNA from Two Plants of Haloxylon Bunge

  • Received:2013-04-15 Revised:2013-05-24 Online:2013-11-25 Published:2013-11-25

摘要: 为了分离提取高质量梭梭(Haloxylon ammodendron)、白梭梭(Haloxylon persicum)线粒体DNA,以藜科梭梭属植物梭梭、白梭梭黄化苗为材料,通过采用蔗糖密度梯度离心和差速离心相结合的方法,使用DNaseⅠ处理得到了无核DNA污染的线粒体,用3% CTAB裂解线粒体,经氯仿/异戊醇抽提除去蛋白,从而得到单纯线粒体DNA。结果表明,所提取的线粒体DNA经电泳检测条带清晰,无拖尾现象,且A260/A280≈1.8,说明纯度较高、质量好,可满足后续有关线粒体DNA的相关实验。梭梭、白梭梭高纯度线粒体DNA提取技术的建立,为开展梭梭、白梭梭线粒体基因组研究奠定了基础。

关键词: 残留动态, 残留动态

Abstract: In order to extract mitochondrial DNA of high-quality Haloxylon ammodendron and Haloxylon persicum, the etiolated seedlings of Haloxylon ammodendron and Haloxylon persicum were taken as experimental materials. This research had established a method of Nuclear-free DNA mitochondrion was firstly obtained by combining sucrose density gradient centrifugation and differential centrifugation together and using DNaseⅠ. Then after the mitochondrion was lysed by using 3% CTAB and its protein was removed by chloroform-isoamyl alcohol extraction, purified mitochondrial DNA was finally got. With A260/A280≈1.8, the result showed that the extracted DNA presented distinct bands and no streaking during the electrophoresis, demonstrating that it enjoys high quality and purity, and can be applied in further experiments concerning mitochondrion DNA. The establishment of this method has laid a foundation for study on mitochondrial genome of Haloxylon ammodendron and Haloxylon persicum.