梭梭属2种植物线粒体基因组的提取-中国农学通报.pdfVIP

中国农学通报 2013,29(33):319-322 Chinese Agricultural Science Bulletin 梭梭属2 种植物线粒体基因组的提取 1,2 1,2 1,2 1 1 1 吴明朝 ，张明慧 ，杨丽丽 ，石 磊 ，陈虞超 ，宋玉霞 1 2 （宁夏农业生物技术重点实验室，银川750002 ；宁夏大学生命科学学院，银川750021） 摘 要：为了分离提取高质量梭梭(Haloxylon ammodendron)、白梭梭(Haloxylon p ersicum) 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DNase Ⅰ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：梭梭；白梭梭；线粒体；线粒体基因组；提取 中图分类号：Q52 文献标志码：A 论文编号：2013-1064 The Study on Extracting Mitochondrial DNA from Two Plants of Haloxylon Bunge 1,2 1,2 1,2 1 1 1 Wu Mingzhao , Zhang Minghui , Yang Lili , Shi Lei , Chen Yuchao , Song Yuxia 1 ( NingxiaAcademyofAgricultureandForestrySciences, Yinchuan 750002; 2 CollegeofLifeScience,NingxiaUniversity, Yinchuan 750021) 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 DNase Ⅰ. Then after the mitochondrion was lysed by using 3% CTAB and its p