两种从陈旧血中抽提基因组DNA方法的比较  

A comparison study of two methods for genomic DNA extraction from obsolete whole blood

作　　者：唐斯[1] 王大明[1] 李桢[1] 高素青[1] 邓志辉[1] 

机构地区：[1]广东省深圳市血液中心输血医学研究所,518035

出　　处：《江西医学检验》2007年第5期423-425,共3页Jiangxi Journal of Medical Laboratory Sciences

摘　　要：目的比较盐析法和磁珠法从陈旧血中抽提基因组DNA的效果和特点,以便常规用于骨髓库HLA基因分型。方法使用TECAN全自动工作站为平台,分别用Gentra盐析法DNA抽提试剂盒和Promega磁珠法DNA抽提试剂,从48份陈旧全血样本中抽提基因组DNA,提取的基因组DNA均溶于100μlTE中;然后用紫外分光光度计测定其产量和纯度,并用琼脂糖凝胶电泳法测定DNA样本的完整性。结果从400μl陈旧全血中提取基因组DNA,用盐析法获得的DNA含量为28.6±8.7ng/μl,A260/A280值平均为1.63±0.209;用磁珠法获得的DNA含量为94.2±10.3ng/μl,A260-A320/A280-A320值平均为1.821±0.102;琼脂糖电泳法检测表明两种方法提取的DNA的分子量均大于15kb。结论使用磁珠法从陈旧血中所获得的DNA的产量和纯度明显高于盐析法,适合于骨髓资料库陈旧血样本的常规HLA基因分型实验以及下游的分子生物学实验。Objective To utilize the optimum DNA extraction method in routine HLA genotyping of bone marrow donor program by comparing the yield and quality of genomic DNA extracted from obsolete whole blood samples by the magnetic beads method and the salt out method. Methods Based on the TECAN DNA workstation platform, the genomic DNA of 48 obsolete whole blood samples were extracted automatically by Promega magnetic beads method and Gentra salt out method, respectively; the extracted genomic DNA of each sample was finally dissolve in 100μl TE solution. The concentration and purity of each DNA sample was tested by UV-spectrophotometer, the integrality of these DNA sample were run electrophoresis by agarose gel. Results The mean concentration and purity （A260/A280） of genomic DNA extracted by Gentra salt out method were 28.6±8.7 ng/μl and 1.63±0.209, respectively; the mean concentration and purity （A260-A320/A280-A320） of genomic DNA extracted by Promega magnetic beads method were 94.2±10.3 ng/ul and 1.821±0.102, respectively. The genomic DNA extracted by two methods were run by electrophoresis on the agarose gel and indicated that the molecular weight for 48 DNA samples were all 〉15kb in length. Conclusions Our results indicate that the yield and purity of genomic DNA extracted by magnetic beads method are better than those of GentraTM salt out method, genomic DNA extracted from the obsolete whole blood by magnetic beads method is suitable for routine HLA typing and other downstream molecular biological experiments.

关 键 词：陈旧血 盐析法 磁珠法 全自动工作站 基因组DNA 

分 类 号：R392-33[医药卫生—免疫学]