Assessing Sample Preparation Methods for Emerging DNA Sequencing Technologies in Human Forensic mtDNA Analysis Applications

• NCJ Number: 252006
• Date Published: October 2016
• Author(s): Brittania J. Bintz
• Length: 92 pages
• Grant Number(s): 2013-DN-BX-K014
• Agencies: NIJ-Sponsored
• Publication Type: Research (Applied/Empirical); Program/Project Evaluation

Annotation

Findings and methodology are presented for a research project whose ultimate goal was to continue to develop methods that enable the generation of whole mt-genome DNA sequence information from compromised or limited DNA samples, so as to expand the potential use of this marker system.

Abstract

This project developed working protocols that capture the entire mtGenome sequence at sufficient depth to identify and compare variants between forensic samples such as blood, buccal scrapes, and hair. This research has demonstrated that whole mtGenome information may be obtained from compromised human samples that include hair shafts and calcified tissues. In order to obtain whole mtDNA genome information from hair shaft material, researchers used enzymatic pre-amplification steps known as whole genome amplification, multiplexed PCR amplification of targeted mtDNA regions, a simple enzymatic library preparation method that uses a transposase-mediated method, followed by 'next generation' DNA sequencing (NGS) of the templates. Further work is warranted in a number of areas related to NGS sequencing in support of forensic casework, including further protocol development, quality-filtering, software package evaluation, advanced mixture studies, validation, and rapid population database creation of the whole mt-genome to support casework analyses. 34 tables, 23 figures, and extensive references under various subject categories