摘要
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.
出处
《生物技术通讯》
CAS
2007年第1期95-97,共3页
Letters in Biotechnology
基金
山东省自然科学基金项目(Y2006D03)
聊城大学实验技术研究项目
关键词
动物
线粒体DNA
分离纯化
animal
mitochondrial DNA
isolation and purification