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用于性侵案件的基于过滤的DNA制备方法。

Filtration based DNA preparation for sexual assault cases.

作者信息

Garvin Alex M

机构信息

Bureco Corporation, 18 Gewerbestrasse, CH-4123 Allschwil, Switzerland.

出版信息

J Forensic Sci. 2003 Sep;48(5):1084-7.

Abstract

Police departments in the United States currently have as many as 500,000 unprocessed swabs taken from rape victims. The standard method for purifying sperm from these swabs is to resuspend first all cells and to digest selectively the excess of the victim's epithelial cells. The intact sperm are then separated from the contaminating solubilized DNA by centrifugation, careful removal of supernatant, and extensive washing of the sperm pellet, all steps that are difficult to automate. Vacuum driven filtration is an alternative method for separating sperm from digested epithelial cells that requires only pipetting steps and can be readily automated in a 96 well format. Sperm DNA is enriched 45-fold using this process and the yield of PCR ready DNA is roughly 20% of the amount originally present on the swab.

摘要

美国各警察局目前有多达50万份从强奸受害者身上采集的未经处理的拭子样本。从这些拭子中纯化精子的标准方法是首先重悬所有细胞,然后选择性地消化掉过量的受害者上皮细胞。然后通过离心、小心去除上清液以及对精子沉淀进行大量洗涤,将完整的精子与污染的溶解DNA分离,所有这些步骤都难以实现自动化。真空驱动过滤是一种从消化后的上皮细胞中分离精子的替代方法,该方法只需要移液步骤,并且可以很容易地以96孔板形式实现自动化。使用该方法,精子DNA可富集45倍,并且可用于PCR的DNA产量约为拭子上最初存在量的20%。

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