单根无毛囊犬毛的自动化 DNA 提取用于线粒体 DNA 分析。

Automated DNA extraction of single dog hairs without roots for mitochondrial DNA analysis.

机构信息

Laboratory of Forensic Genetics and Molecular Archaeology, UZ Leuven, Kapucijnenvoer 33, B-3000 Leuven, Belgium.

出版信息

Forensic Sci Int Genet. 2012 Mar;6(2):277-81. doi: 10.1016/j.fsigen.2011.04.009. Epub 2011 Apr 29.

DOI:10.1016/j.fsigen.2011.04.009

Abstract

Dogs are intensely integrated in human social life and their shed hairs can play a major role in forensic investigations. The overall aim of this study was to validate a semi-automated extraction method for mitochondrial DNA analysis of telogenic dog hairs. Extracted DNA was amplified with a 95% success rate from 43 samples using two new experimental designs in which the mitochondrial control region was amplified as a single large (± 1260 bp) amplicon or as two individual amplicons (HV1 and HV2; ± 650 and 350 bp) with tailed-primers. The results prove that the extraction of dog hair mitochondrial DNA can easily be automated to provide sufficient DNA yield for the amplification of a forensically useful long mitochondrial DNA fragment or alternatively two short fragments with minimal loss of sequence in case of degraded samples.

摘要

狗在人类的社会生活中占据着重要地位，它们的脱落毛发在法医调查中起着重要作用。本研究的总体目标是验证一种用于分析休止期狗毛的线粒体 DNA 的半自动提取方法。使用两种新的实验设计，从 43 个样本中以 95%的成功率扩增出提取的 DNA，其中线粒体控制区被扩增为单个大的（约 1260 bp）扩增子，或作为两个带有长尾引物的单独扩增子（HV1 和 HV2；约 650 和 350 bp）。结果证明，狗毛线粒体 DNA 的提取可以很容易地自动化，从而为扩增具有法医用途的长线粒体 DNA 片段或在降解样本的情况下为扩增两个短片段提供足够的 DNA 产量，而不会损失最小的序列。