DNA数据库建设中批量样品不同DNA提取方法的比较

目的比较和选择自动化工作平台提取DNA的方法,并用于DNA数据库建设。方法用手工Chelex-100法、Biomek3000自动化工作平台结合Chelex-100法及DNA-IQTM磁珠法对实验室收集的建库滤纸血样进行DNA提取,荧光定量技术对上述3种方法提取的模板DNA进行测定;扩增产物用3100基因分析仪检测并用基因分析软件分析。结果手工Chelex-100法、自动化Chelex-100法及DNA-IQTM磁珠法提取的DNA模板浓度分别为0.593ng±0.131ng/μl、0.579ng±0.096ng/μl、0.447ng±0.056ng/μl;成功率分别为100%、98.9%、99.5%。结论本文建立的自动化Chelex-100法可用于大规模DNA数据库建设。

Comparison on different DNA extraction methods for batches of bloodstain samples during the establishment of national DNA database

Objective To compare and evaluate different DNA extraction methods for batches of bloodstain samples during the establishment of national DNA database. MethodsDNA was extracted from bloodstain samples collected for the establishment of DNA database using manual Chelex-100, Biomek 3000 automation workstation with Chelex-100 and DNA-IQTM magnetic bead method respectively, and the quantity of DNA was determined by the real-time fluorescent quantitative PCR technique. Genotyping was performed using the AmpFLSTR STR Identifiler kit and 3100 genetic analyzer. ResultsThe concentrations of DNA templates obtained by manual Chelex-100, automatic Chelex-100 and DNA-IQTM magnetic bead method were 0.593±0.131ng/μl, 0.579±0.096ng/μl and 0.447±0.056ng/μl respectively, and the success rates of genotyping with these DNA templates were 100%, 98.9% and 99.5% respectively. ConclusionThe automatic Chelex-100 method proposed by this study can be applied in the establishment of large-scale DNA databases.