Rapid direct PCR for forensic genotyping in under 25 min
Rapid direct PCR for forensic genotyping in under 25 min
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DOI：
10.1002/elps.201200570
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发表时间：
2013-06-01
期刊：
影响因子：
2.9
通讯作者：
McCord, Bruce
中科院分区：
文献类型：
--
作者：
Aboud, Maurice;Oh, Hye Hyun;McCord, Bruce
在本文中，快速热循环程序与打孔机的直接扩增相结合，允许高速扩增 7 基因座多重信号，无需提取步骤。当与短的 1.8 cm 微流体电泳系统结合使用时，从打孔到基因分型的整个过程可以在 25 分钟内完成。本文描述了酶的选择和优化、直接扩增条件、程序的重现性以及与标准法医基因分型方法的一致性。该程序使用小型高速热循环仪和微流体装置以及小型笔记本电脑，并且高度便携。总体而言，该技术应为快速确定检查站保留的个人身份提供有用且可靠的程序，并为大规模灾难后在偏远地区初步识别个人提供快速方法。
In this paper, a rapid thermal cycling procedure is combined with a direct amplification from a paper punch, permitting a high-speed amplification of a 7-locus multiplex that requires no extraction step. When coupled with a short 1.8 cm microfluidic electrophoresis system, the entire procedure from paper punch to genotype can be completed in under 25 min. The paper describes selection and optimization of enzyme, direct amplification conditions, the reproducibility of the procedure, and concordance with standard forensic genotyping methods. The procedure utilizes a small high-speed thermal cycler and microfluidic device along with a small laptop and is highly portable. Overall, this technique should provide a useful and reliable procedure for rapid determination of identity of individuals retained at checkpoints as well as a quick method for preliminary identification of individuals at remote locations following mass disasters.