一种改进的提取人外周血和组织线粒体DNA的方法

An improved method for mitochondrial DNA preparation from human peripheral blood and tissues

目的 :建立一种简便、快捷地从人外周血和组织中制备高质量线粒体 DNA的方法。 方法 :以差速离心法分离线粒体 ,用碱性 SDS法裂解线粒体膜 ,释放线粒体 DNA后用酚 /氯仿抽提 ,TE或 dd H2 O溶解。然后将所得线粒体 DNA进行纯度鉴定并和其他现有方法制备的线粒体 DNA用 PCR进行长片段扩增 ,比较扩增效果。结果 :用改进的方法制备的线粒体 DNA中没有检测到核 DNA,并能有效扩增 8kb长的目的片段。 结论 :改进后的方法简便、快捷 ,制备的线粒体 DNA纯度高 ,也有利于长片段

Objective:To develop a simple and rapid method to isolate high quality mitchondrial DNA (mtDNA) from human peripheral blood and tissues.Methods:Mitochondria were isolated by velocity gradient centrifugation and the mitochondrion membrane was broken up by alkaline lysis.mtDNA was then extracted by phonl and chlorodorm.After identification the mtDNA was compared with mtDNAs obtained with routine methods by PCR amplification.Results:The nuclear DNA was not detected in mtDNA prepared by improved method and a 8 kb product was obtained when using it as long PCR template.Conclusion:It is suggested that our method is simple and rapid for preparing mtDNAs with high purity,which is easy for PCR amplification.