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化学和热浸解技术对从骨骼中回收核DNA和线粒体DNA的影响。

The effects of chemical and heat maceration techniques on the recovery of nuclear and mitochondrial DNA from bone.

作者信息

Steadman Dawnie Wolfe, DiAntonio Lisa L, Wilson Jeremy J, Sheridan Kevin E, Tammariello Steven P

机构信息

Department of Anthropology, Binghamton University, SUNY, PO Box 6000, Binghamton, NY 13902-6000, USA.

出版信息

J Forensic Sci. 2006 Jan;51(1):11-7. doi: 10.1111/j.1556-4029.2005.00001.x.

DOI:10.1111/j.1556-4029.2005.00001.x
PMID:16423217
Abstract

Forensic anthropologists use a number of maceration techniques to facilitate skeletal analysis of personal identity and trauma, but they may unwittingly eliminate valuable DNA evidence in the process. This study evaluated the effect of 10 maceration methods on gross bone structure and the preservation of DNA in ribs of 12 pigs (Sus scrofa). A scoring system was applied to evaluate the ease of maceration and resulting bone quality while DNA purity was quantified by optical densitometry analysis, followed by polymerase chain reaction (PCR) amplification of three mitochondrial and three nuclear loci. The results demonstrated that while mitochondrial DNA could be amplified for all experiments, cleaning treatments using bleach, hydrogen peroxide, ethylenediaminetetraacetic acid/papain, room temperature water and detergent/sodium carbonate followed by degreasing had low DNA concentrations and failed to generate nuclear PCR products. In general, treatments performed at high temperatures (90 degrees C or above) for short durations performed best. This study shows that traditionally "conservative" maceration techniques are not necessarily the best methods to yield DNA from skeletal tissue.

摘要

法医人类学家使用多种浸软技术来促进对个人身份和创伤的骨骼分析,但他们可能在这个过程中不经意地消除了有价值的DNA证据。本研究评估了10种浸软方法对12头猪(野猪)肋骨的大体骨骼结构和DNA保存的影响。应用评分系统来评估浸软的难易程度和所得骨骼质量,同时通过光密度分析对DNA纯度进行定量,随后对三个线粒体基因座和三个核基因座进行聚合酶链反应(PCR)扩增。结果表明,虽然所有实验都能扩增出线粒体DNA,但使用漂白剂、过氧化氢、乙二胺四乙酸/木瓜蛋白酶、室温下水和洗涤剂/碳酸钠进行清洁处理,随后脱脂,DNA浓度较低,未能产生核PCR产物。一般来说,在高温(90摄氏度或以上)下进行短时间的处理效果最佳。这项研究表明,传统上“保守”的浸软技术不一定是从骨骼组织中获取DNA的最佳方法。

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