摘要
以油菜为实验材料,探讨了植物线粒体DNA提取纯化过程中的影响因素,构建了一套快捷、经济、高效的植物线粒体DNA提取纯化方法.该方法结合差速离心和密度梯度离心分离得到线粒体,酶消化处理去除其他基因组的污染,用SDS与蛋白酶裂解线粒体,通过高盐沉淀,有机溶剂抽提,最后再用CTAB纯化得到mtDNA.用该方法提取的油菜线粒体DNA,经紫外分光光度计测定其纯度、限制性内切酶进行酶切.结果表明,此提取方法得到的mtDNA,其纯度可以满足后续遗传学分析.该法适用于不同植物材料mtDNA的提取纯化.
Using cole seedlings as experimental materials, this paper made a study of the influencing factors on the isolation and purification of plant mitochondrial DNA. Being regarded as a rapid, economical and efficient method, its protocol includes： isolation of mitochondria by combination of differential eentrifugation and density gradient ultracentrifugation, lysis with SDS and proteinase K, purification by high ionic strength precipitation, organic solvent extraction and CTAB/NaC1 treatment. Tested by UV- speetrophotometer, the mitochondria[ DNA from cole by this protocol is very pure and can be well digested by the restriction endonuclease, and its quality may well enable it to be used in other genetic studies. The protocol is suitable for the extraction of mitochondrial DNA from most plants.
出处
《中南林业科技大学学报》
CAS
CSCD
北大核心
2009年第3期106-110,共5页
Journal of Central South University of Forestry & Technology
基金
湖南省教育厅学科带头人培养对象基金资助项目(2004559)
关键词
生物科学与技术
油菜
线粒体
MTDNA
提取纯化
方法
biological science and technology
cole
mitochondria
mtDNA
isolation and purification
method
