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比较三种不同程度降解的人骨 DNA 提取方法。

Comparison of three methods of DNA extraction from human bones with different degrees of degradation.

机构信息

Intercollegiate Faculty of Biotechnology of University of Gdańsk and Medical University of Gdańsk, Gdańsk, Poland.

出版信息

Int J Legal Med. 2012 Jan;126(1):173-8. doi: 10.1007/s00414-011-0590-5. Epub 2011 Jun 30.

Abstract

There is a necessity for deceased identification as a result of many accidents and sometimes bones are the only accessible source of DNA. So far, a universal method that allows for extraction of DNA from materials at different stages of degradation does not exist. The aims of this study were: the comparison of three methods of DNA extraction from bones with different degree of degradation and an evaluation of the usefulness of these methods in forensic genetics. The efficiency of DNA extraction, the degree of extract contamination by polymerase chain reaction (PCR) inhibitors and the possibility of determining the STR loci profile were especially being compared. Nuclear DNA from bones at different states of degradation was isolated using three methods: classical, organic phenol-chloroform extraction, DNA extraction from crystal aggregates and extraction by total demineralisation. Total demineralisation is the best method for most cases of DNA extraction from bones, although it does not provide pure DNA. DNA extraction from aggregates removes inhibitors much better and is also a good method of choice when identity determination of exhumed remains is necessary. In the case of not buried bones (remains found outside) total demineralisation or phenol-chloroform protocols are more efficient for successful DNA extraction.

摘要

由于许多事故的发生,对死者进行身份识别是必要的,而有时骨骼是唯一可获取 DNA 的来源。到目前为止,还没有一种通用的方法可以从不同降解阶段的材料中提取 DNA。本研究的目的是:比较从不同降解程度的骨骼中提取 DNA 的三种方法,并评估这些方法在法医遗传学中的有用性。特别比较了 DNA 提取效率、聚合酶链反应(PCR)抑制剂污染程度以及确定 STR 基因座图谱的可能性。使用三种方法从不同降解状态的骨骼中分离核 DNA:经典方法、有机酚-氯仿提取法、从晶体聚集体中提取 DNA 以及完全脱矿提取法。对于大多数情况下从骨骼中提取 DNA,完全脱矿是最佳方法,尽管它不能提供纯 DNA。从聚集体中提取 DNA 可以更好地去除抑制剂,而且在需要确定挖掘出的遗骸的身份时,也是一种很好的选择方法。在未埋葬骨骼(在外部发现的遗骸)的情况下,完全脱矿或酚-氯仿方案更有利于成功提取 DNA。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9183/3254873/96e6c6cadeda/414_2011_590_Fig1_HTML.jpg

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