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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
PMID:21531187
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。

相似文献

1
Automated DNA extraction of single dog hairs without roots for mitochondrial DNA analysis.单根无毛囊犬毛的自动化 DNA 提取用于线粒体 DNA 分析。
Forensic Sci Int Genet. 2012 Mar;6(2):277-81. doi: 10.1016/j.fsigen.2011.04.009. Epub 2011 Apr 29.
2
A simplified method for mitochondrial DNA extraction from head hair shafts.一种从头发毛囊线粒体DNA提取的简化方法。
J Forensic Sci. 2005 Sep;50(5):1119-22.
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Identification of forensically informative SNPs in the domestic dog mitochondrial control region.家犬线粒体控制区法医信息性单核苷酸多态性的鉴定
J Forensic Sci. 2009 Mar;54(2):289-304. doi: 10.1111/j.1556-4029.2008.00953.x.
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Forensic DNA-typing of dog hair: DNA-extraction and PCR amplification.狗毛的法医DNA分型:DNA提取与聚合酶链反应扩增
Forensic Sci Int. 2004 May 10;141(2-3):149-51. doi: 10.1016/j.forsciint.2004.01.016.
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Mitochondrial DNA heteroplasmy among hairs from single individuals.单一个体毛发中的线粒体DNA异质性
J Forensic Sci. 2004 Sep;49(5):986-91.
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A novel method for forensic DNA investigations: repeat-type sequence analysis of tandemly repeated mtDNA in domestic dogs.一种用于法医DNA调查的新方法:家犬串联重复线粒体DNA的重复类型序列分析
J Forensic Sci. 2000 Sep;45(5):990-9.
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Sequence analysis of domestic dog mitochondrial DNA for forensic use.用于法医鉴定的家犬线粒体DNA序列分析。
J Forensic Sci. 1997 Jul;42(4):593-600.
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Usefulness of microchip electrophoresis for the analysis of mitochondrial DNA in forensic and ancient DNA studies.微芯片电泳在法医和古DNA研究中线粒体DNA分析中的应用价值。
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Sequence analysis of the canine mitochondrial DNA control region from shed hair samples in criminal investigations.刑事调查中脱落毛发样本的犬线粒体DNA控制区序列分析。
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[Mitochondrial DNA sequencing of human hair shafts stored for long time].[长期保存的人类毛发线粒体DNA测序]
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引用本文的文献

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Animals (Basel). 2023 Mar 12;13(6):1036. doi: 10.3390/ani13061036.
2
Who killed my dog? Use of forensic genetics to investigate an enigmatic case.谁杀了我的狗?利用法医遗传学调查一个神秘案件。
Int J Legal Med. 2021 Mar;135(2):387-392. doi: 10.1007/s00414-020-02388-9. Epub 2020 Aug 11.
3
Mitochondrial diversity of Bulgarian native dogs suggests dual phylogenetic origin.保加利亚本土犬的线粒体多样性表明其具有双重系统发育起源。
PeerJ. 2018 Jun 27;6:e5060. doi: 10.7717/peerj.5060. eCollection 2018.
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Reviewing population studies for forensic purposes: Dog mitochondrial DNA.出于法医目的回顾群体研究:犬线粒体DNA
Zookeys. 2013 Dec 30(365):381-411. doi: 10.3897/zookeys.365.5859.
5
Successful direct amplification of nuclear markers from a single hair follicle.从单个毛囊成功直接扩增核标记物。
Forensic Sci Med Pathol. 2013 Jun;9(2):238-43. doi: 10.1007/s12024-012-9402-6. Epub 2012 Dec 30.