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应用生物系统公司Prism 377型自动测序仪用于法医短串联重复序列分析的验证

Validation of the Applied Biosystems Prism 377 automated sequencer for the forensic short tandem repeat analysis.

作者信息

Frazier R R, Millican E S, Watson S K, Oldroyd N J, Sparkles R L, Taylor K M, Panchal S, Bark L, Kimpton C P, Gill P D

机构信息

Forensic Science Service, Birmingham, UK.

出版信息

Electrophoresis. 1996 Oct;17(10):1550-2. doi: 10.1002/elps.1150171008.

Abstract

The Applied Biosystems (ABI) Prism 377 DNA sequencer has been evaluated in an attempt to increase the throughput of samples for short tandem repeat (STR) analysis, in both forensic casework and the UK National Criminal Intelligence DNA Database. The gel system assessed consisted of 0.2 mm, 4% acrylamide 6 M urea gels, with a well-to-read distance of 36 cm. Gels were run at a constant voltage of 3 kV and constant temperature of 51 degrees C. The run time of our second generation multiplex (SGM) STR system was achieved in less than 2 h. Rigorous validation has been performed on the instrument hardware and software. Complete resolution of 1 base differences was obtained, up to and beyond 350 bases; sizing precision across gels was more than 2-fold higher than the 373A and the sensitivity was increased by one third.

摘要

应用生物系统公司(ABI)的377型DNA测序仪已通过评估,旨在提高法医案件工作和英国国家刑事情报DNA数据库中短串联重复序列(STR)分析的样本通量。所评估的凝胶系统由0.2毫米、4%丙烯酰胺6M尿素凝胶组成,孔到读数的距离为36厘米。凝胶在3千伏的恒定电压和51摄氏度的恒定温度下运行。我们的第二代复合扩增(SGM)STR系统的运行时间不到2小时。已对仪器硬件和软件进行了严格验证。获得了高达350个碱基及以上的1个碱基差异的完全分辨率;凝胶间的大小测定精度比373A型高出两倍多,灵敏度提高了三分之一。

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