动物线粒体DNA提取简易流程及其优化

Study on Method for Isolation of Mithochondrial DNA in Animal

目的:研究动物线粒体DNA(mtDNA)提取的简易流程,以提取到纯度高、结构完整的动物mtDNA。方法:采用SDS碱变性法,从动物的肝脏和性腺等组织中提取mtDNA,并增加了DNaseⅠ、RNase消化步骤,以除去核DNA及RNA的污染;用紫外分光光度计分析mtDNA的纯度和得率。结果:mtDNA的得率为0.5～0.8μg/g,D260nm/D280nm值为1.77～1.82,能满足对mtDNA进行RFLP分析、RAPD分析、测序分析等的要求。结论:改进的动物mtDNA提取方法简便可行,值得推广。

Objective： To establish a kind of method used for the isolation and purification of mitochondrial DNA（mtDNA） from animal. Methods： The mtDNA was isolated and purified by the means of SDS alkali decomposition from liver and gonad of animals. The procedure of DNase Ⅰ , RNase digestion was added for avoiding completely the possible pollution with nuclear DNA, RNA, etc. The mtDNA was identified by UV spectrophotometer. Results： The quality of mtDNA obtained was 0.5-0.8μg/g, and the D260nm/D280nm was 1.77-1.82. The mtDNA obtained was suitable to be analyzed by RFLP, RAPD, sequence and other kinds of methods. Conclusion： The method established is more quicker and simpler than former methods, and is worth to popularize.