Korber Jade T, Canale Lauren C, Holland Mitchell M
Department of Biochemistry & Molecular Biology, Forensic Science Program, The Pennsylvania State University, University Park, Pennsylvania.
Department of Justice, Jan Bashinski DNA Laboratory, Richmond, California.
Curr Protoc. 2023 Aug;3(8):e865. doi: 10.1002/cpz1.865.
This article highlights methods used to perform DNA extraction, mitochondrial DNA quantification, multiplex PCR amplification, amplicon-based massively parallel sequencing, and data analysis of the mitochondrial genome (mitogenome) from human hair shafts. The focus is on applications to forensic casework, but this set of protocols can be used for any purpose involving small cuttings (as small as 1 to 5 mm) of human hair shafts up to 40 years from the time of collection. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Extraction of mitochondrial DNA from human hair shafts Basic Protocol 2: Quantification of mitochondrial DNA (copies/μl) Basic Protocol 3: Multiplex amplification of the mitogenome Basic Protocol 4: Library preparation and sequencing of mitogenome amplicons Basic Protocol 5: Data analysis of mitogenome haplotypes.
本文重点介绍了用于从人类毛发中进行DNA提取、线粒体DNA定量、多重PCR扩增、基于扩增子的大规模平行测序以及线粒体基因组（有丝分裂基因组）数据分析的方法。重点在于法医案件工作中的应用，但这组方案可用于任何涉及从采集时间起长达40年的人类毛发小片段（小至1至5毫米）的目的。© 2023作者。由Wiley Periodicals LLC出版的《当前协议》。基本方案1：从人类毛发中提取线粒体DNA 基本方案2：线粒体DNA定量（拷贝数/微升） 基本方案3：有丝分裂基因组的多重扩增 基本方案4：有丝分裂基因组扩增子的文库制备和测序 基本方案5：有丝分裂基因组单倍型的数据分析。
