艾滋病患者外周血单个核细胞DNA的提取方法

DNAextractionfromperipheralbloodmononuclearcelinpatientswithAIDS.FanXueli,FanJiang,XuWenyan,etal.InstituteofDermatology,ChineseAcademyofMedicalScience.Nanjing,210042ObjectivePrepareingthehighqualityDNAfromperipheralbloodmononuclearcelinpatientswithAIDSorH

目的：为了完整地获得艾滋病患者或感染者外周血单个核细胞中的ＤＮＡ分子，以适应其分子流行病学研究的需要。方法：分离外周血单个核细胞，采用尿素－十二烷基硫酸钠裂解缓冲液破碎细胞，经酚、氯仿、异戊醇等抽提蛋白、乙醇沉淀核酸等步骤提取ＤＮＡ样品。结果：样品中ＤＮＡ含量较高，结构稳定，聚合酶链反应扩增效果良好。结论：本法提取的ＤＮＡ样品适合于基因扩增和分子克隆。

Objective Prepareing the high quality DNA from peripheral blood mononuclear cell in patients with AIDS or HIV infection,to satisfy the study on molecular epidemiology of AIDS.Method The peripheral blood mononuclear cells isolated from AIDS patients were digested with Urea SDS lysis buffer.The DNA samples were extracted with phenol,chloroform and isopentanol.The final DNA was prepared after we rinsed the pellet of DNA with ethanol.Result DNA prepared in this way was sufficient and had stable structure.It was confirmed by PCR that the HIV 1 gag gene could be amplified effectively from the PBMC DNA.Conclusion The DNA samples extracted above were suitable for genetic amplification and cloning.It laid a fundation for the early diagnosis and further study of AIDS.