摘要
目的使用Chelex-100方法在核酸载体分离板(96孔)上对接触性生物检材DNA进行载体分离和纯化,并将其与TECAN Freedom EVO150-8自动化工作站进行结合,建立起一种自动化、快速、灵敏、高效的DNA提取方法并探讨其在法医学中的应用价值。方法在核酸载体分离板(96孔)上使用Chelex-100液裂解DNA,通过高速离心实现批量生物检材载体与DNA分离;通过编写EVO150-8自动化工作站应用程序实现批量生物检材的快速提取、PCR构建一体化过程;采用。Identifiler Plus试剂盒进行PCR扩增,扩增产物经3500 xL遗传分析仪电泳,GeneMapperID-X软件分析检测结果。结果在994份生物检材中,成功获得456份检材STR分型,平均检验成功率在45.88%;对92份生物检材DNA提取及PCR构建,仅耗时60 min。结论本研究应用核酸载体分离板(96孔),建立的自动化Chelex-100法快速、高效、灵敏,适用于批量接触性检材的DNA检测。
Objective To establish a batch extraction method for touch DNA by Chelex-100 injection into 96-well nucleic acid filtration plate through TECAN Freedom EVO150-8 automation workstation so as to build up an automatic, rapid, sensitive and highly efficient way to isolate and purify DNA. Methods Biological samples were lysed by Chelex-100 infusing into 96-well lysis purification plate. The lysate DNA was isolated from the carrier and precipitated into 96-well centrifugal collecting plate by high speed centrifugation. Automatic application programs were written for TECAN Freedom EVO 150-8 to achieve both the rapid DNA extraction from massive biological samples and PCR construction. Amplified by Identifiler Plus kit, the DNA template and its product were subjected to capillary electrophoresis in 3500XL analyzer, having the resultant data analyzed with GeneMapper ID-X software. Results Of all 994 samples, 456 ones were successfully detected of their STR genotypes, obtaining a success rate of 45.88% for the detection of touch DNA. It takes only 58 minutes for 92 biological samples to conduct the DNA extraction and PCR construction with the method established here. Conclusions The DNA extraction method, that of isolating and purifying DNA by chelex-100 injecting into 96-well filtration plate through TECAN Freedom EVO 150-8 automation workstation, is rapid, efficient and sensitive, applicable for massive touch DNA detection.
出处
《刑事技术》
2017年第3期206-208,共3页
Forensic Science and Technology
基金
江苏省公安厅科研项目(No.201101001)
