Recovery of Whole Mitochondrial Genome From Compromised Samples via Multiplex PCR and Massively Parallel Sequencing

• NCJ Number: 254295
• Author(s): Maureen P. Hickman; Kelly S. Grisdale; Brittania J Bintz; Erin S. Burnside; Erin K. Henson; Jack ballantyne; Mark R. Wilson
• Date Published: 2018
• Length: 12 pages
• Grant Number(s): 2013-DN-BX-K014
• Agencies: NIJ-Sponsored
• Publication Type: Research (Applied/Empirical); Report (Study/Research); Report (Grant Sponsored); Program/Project Description
• Award: Assessing Sample Preparation Methods for Emerging DNA Sequencing Technologies in Human Forensic mtDNA Analysis Applications

Annotation

This article reports on a project that developed a multiplex PCR method that amplifies the complete mitochondrial genome in a range of forensically relevant samples, including single cells, cremated remains, bone, maggot, and hairs isolated from dust bunnies.

Abstract

In forensic casework, compromised samples often have limited or degraded nuclear DNA, rendering mitochondrial DNA a more feasible option for forensic DNA analyses. The emergence of massively parallel sequencing (MPS) has enabled the recovery of extensive sequence information from low quantities of DNA. In the current project, MPS yields completed or nearly completed mitochondrial genome coverage for all samples. To confirm concordance between sample types and between sequencing platforms, the project compared sequencing results from hair and buccal swabs from two references. Low initial DNA input into the multiplex PCR allowed for conservation of precious DNA, while MPS maximized recovery of genetic information. (publisher abstract modified)