在不脱矿的情况下从骨骼中恢复DNA

IF 0.5 Q4 GENETICS & HEREDITY
Sasitaran Iyavoo , William H. Goodwin
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引用次数: 1

摘要

本研究评估了从骨骼中回收DNA的五种不同DNA提取方法的性能:ChargeSwitch®gDNA植物试剂盒、DNA IQ™ 系统套件,DNeasy®Blood&;纸巾试剂盒、PrepFiler®BTA法医DNA提取试剂盒和苯酚-氯仿-异戊醇。从新鲜的猪肋骨和股骨中提取DNA,这些骨头经过了三个月的表面分解,并且经过了一年的表面分解。使用实时PCR和内部PCR多重扩增分析提取的DNA,该多重扩增评估DNA的质量和数量以及抑制剂的存在。基于苯酚-氯仿的方法始终产生最高量的DNA,DNA IQ最低;然而,所有方法都从猪肋骨和股骨样本中产生了相对较高产量的DNA,这些DNA可以在没有任何检测到的抑制的情况下进行扩增。数据表明,只要骨骼样本质量合理,任何测试方法都可以分离出可以成功分析的DNA。还证明了内部PCR对照的有效使用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Recovery of DNA from bone without demineralization

This study assessed the performance of five different DNA extraction methods for the recovery of DNA from bone: ChargeSwitch® gDNA Plant Kit, DNA IQ™ System Kit, DNeasy® Blood & Tissue Kit, PrepFiler® BTA Forensic DNA Extraction Kit and phenol-chloroform-isoamyl alcohol. DNA was extracted from pig rib and femur bones that was fresh, had undergone surface decomposition for three months, and had undergone surface decomposition for one year. Extracted DNA was analyzed using real-time PCR and amplification of an in-house PCR multiplex that assessed the quality and quantity of DNA and for the presence of inhibitors. The phenol-chloroform-based method consistently yielded the highest amounts of DNA and DNA IQ the lowest; however, all methods produced relatively high yields of DNA from both pig rib and femur samples that could be amplified without any detected inhibition. The data demonstrate that with reasonable quality bone samples any of the tested methods can isolate DNA that can be successfully analyzed. The effective use of internal PCR controls is also demonstrated.

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来源期刊
Forensic Science International: Genetics Supplement Series
Forensic Science International: Genetics Supplement Series Medicine-Pathology and Forensic Medicine
CiteScore
0.40
自引率
0.00%
发文量
122
审稿时长
25 days
期刊介绍: The Journal of Forensic Science International Genetics Supplement Series is the perfect publication vehicle for the proceedings of a scientific symposium, commissioned thematic issues, or for disseminating a selection of invited articles. The Forensic Science International Genetics Supplement Series is part of a duo of publications on forensic genetics, published by Elsevier on behalf of the International Society for Forensic Genetics.
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