TE-MAGS磁珠法提取效率定量研究  

Quantification study of TE-MAGS magnetic bead based DNA extraction efficiency

作　　者：韩海军 苏勇[2] 贾东涛 张玉红 朱奇慧 马腾[3] 

机构地区：[1]南通市公安局物证鉴定所,江苏南通226007 [2]江苏省公安厅物证鉴定中心,江苏南京210024 [3]复旦大学生命科学学院,上海200433

出　　处：《中国法医学杂志》2017年第6期614-617,共4页Chinese Journal of Forensic Medicine

基　　金：江苏省公安厅科研重点项目(2016KF018Z)

摘　　要：目的利用TE-MAGS在TECAN自动化工作站上结合磁珠试剂盒,定量研究微量DNA提取回收效率和纯化能力。方法 0.1~1ng9947A和混有6种常见PCR抑制剂的1ng9947A,经自动化提取后进行荧光定量和STR分型,定量分析回收效率和纯化能力。结果 0.1~1ng9947A提取后实际回收效率在38.92~60.01%之间,0.3ng以上9947A提取扩增后可以得到完整的STR图谱。胆汁酸盐、胶原质、尿素去除效率大于94.5%,血红素、黑色素、腐殖酸去除效率分别为97.5%、97.85%、82.14%。结论 TE-MAGS磁珠法对微量DNA回收效率较高,纯化能力较强,适合微量DNA提取。Objective This study aimed to assess the efficiency and purification of the Trace DNA extraction with a quantified method for the magnetic bead-based DNA extraction as performed on the Tecan Automated systems with TE-MAGS magnetic separator.Methods Serial dilutions of standard commercial DNA 9947 A were used with the total DNA contents,0.1 ng,0.2 ng,0.3 ng,0.4 ng,0.5 ng,0.6 ng,0.7 ng,0.8 ng,0.9 ng,1 ng,diluted progressively and a 1 ng DNA（standard commercial DNA 9947 A） admixed with 6 common DNA-PCR inhibitors were extracted on the Automated systems and then performed via Fluorogenic probe quantitative PCR and STR genotype for the quantification analysis of recovery and purification.Results The recovery rate of standard 9947 A DNA diluted with 0.1~1 ng was 38.92~60.01%,and 0.3 ng and more DNA could observed the full STR profiles.For the different PCR inhibitors,above 94.5% of bile acid,collagen and urea were efficient removal during the purification process,and the hemoglobin,melanin and humic acid removal efficiency were about 97.5%,97.85%,82.14%,respectively.Conclusion Our results suggested that The TE-MAGS magnetic bead-based DNA extraction was suitable for the extraction of trace DNA with high recovery efficiency and purification ability.

关 键 词：法医物证学 微量DNA 定量 TE-MAGS磁珠法 自动化DNA提取 

分 类 号：D919[医药卫生—法医学]