摘要
目的为适应大批量提取外周血DNA需求,针对现有试剂盒提取法进行改良。方法将武汉地区健康人群血液样本80份,随机分为两组,每组40份。分别采用改良全血DNA提取法和外周血单个核细胞(peripheral blood mononuclear cell,PBMC)DNA提取法提取DNA,比较其效度和质量。结果通过比较分析实验时间、提取DNA的质量、纯度和PCR结果,发现改良全血DNA提取法提取的时间显著少于全血PBMC DNA提取法,提取DNA质量显著高于全血PBMC DNA提取法(P<0.05),而两种方法提取的基因组DNA的OD260/OD280数值差异无统计学意义(P>0.05),且PCR产物均可见单一且明亮的条带。结论本实验中改良全血DNA提取法是一种快速、经济、高效的DNA提取方法,可在临床移植配型、临床基因多样性检测中发挥作用。
Objective To meet the demand for large-scale extraction of peripheral blood DNA,we make improvements in the existing kit extraction method.Methods Eighty blood samples from healthy people in Wuhan were randomly divided into two groups with 40 samples in each group.Improved whole blood DNA extraction method and PBMC(peripheral blood mononuclear cell)DNA extraction method were used to extract DNA respectively,and their validity and quality were compared.Results By comparing and analyzing the experimental time,the quality and purity of the extracted DNA,and the PCR results,it was found that the improved whole blood DNA extraction method took significantly less time than the whole blood PBMC DNA extraction method,and the extracted DNA quality was significantly higher than the whole blood PBMC DNA extraction method(P<0.05),while the OD260/OD280 values of the genomic DNA extracted by the two methods were not statistically different(P>0.05),and single and bright bands can be seen in the PCR products of the two methods.Conclusion The improved whole blood DNA extraction method is a fast,economical,and efficient DNA extraction method,which can play a role in clinical transplantation matching and clinical genetic diversity detection.
作者
左文博
王嘉政
徐颖
杨小婷
倪维
胡松
ZUO Wenbo;WANG Jiazheng;XU Ying;YANG Xiaoting;NI Wei;HU Song(School of Medicine,Jianghan University,Wuhan 430056,Hubei,China;Hubei Provincial Hospital of TCM,Wuhan 430061,Hubei,China)
出处
《江汉大学学报（自然科学版）》
2020年第5期42-46,共5页
Journal of Jianghan University：Natural Science Edition
基金
国家自然科学基金资助项目(31470270)
江汉大学2018年度学生学术科技项目(2018yb249)。
