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用于低模板DNA样本处理的DNA纯化、细胞裂解及洗涤步骤的改进

DNA Purification Cell Lysis and Wash Step Modifications for Low-Template DNA Sample Processing.

作者信息

Menchhoff Sydney I, Delacruz Milady T, Hytinen Madison E, Cox Jordan O, Miller Marilyn T, Dawson Cruz Tracey

机构信息

Department of Forensic Science, Virginia Commonwealth University, 1015 Floyd Avenue, Room 2015, Richmond, VA, 23284.

出版信息

J Forensic Sci. 2020 Mar;65(2):597-600. doi: 10.1111/1556-4029.14203. Epub 2019 Oct 2.

Abstract

As DNA technology becomes increasingly sensitive, forensic laboratories are receiving more low-template DNA samples. These samples, already low in DNA content, become even more challenging to process as the available DNA becomes further reduced during the extraction step. In this study, two extraction modifications were tested to determine if the cause of DNA loss could be identified and mitigated. A double lysis technique was used to test for DNA loss in the sample collection substrate, and lysate eluates were re-extracted to determine DNA loss from inefficient binding to the silica column. Both modifications showed DNA was lost at these steps. However, resulting STR profiles from these samples had fewer peaks and lower peak heights when compared to samples processed with no extraction modifications. Overall, the potential benefits of adding these extraction modifications for low-template DNA sample processing are not enough to justify the risk associated with additional manipulation.

摘要

随着DNA技术变得越来越灵敏,法医实验室收到的低模板DNA样本越来越多。这些样本本身DNA含量就低,而在提取步骤中可用DNA进一步减少时,处理起来就更具挑战性。在本研究中,测试了两种提取方法的改进,以确定是否能够识别并减轻DNA损失的原因。采用双重裂解技术来测试样本采集底物中的DNA损失,对裂解液洗脱物进行重新提取,以确定因与硅胶柱结合效率低下而导致的DNA损失。两种改进方法均显示在这些步骤中存在DNA损失。然而,与未进行提取方法改进处理的样本相比,这些样本得到的STR图谱峰数更少、峰高更低。总体而言,对低模板DNA样本处理采用这些提取方法改进的潜在益处不足以证明与额外操作相关的风险是合理的。

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