NCJ Number:
247278
Title:
Assessing Deep Sequencing Technology for Human Forensic Mitochondrial DNA Analysis
Author(s):
Mark R. Wilson, Ph.D.
Date Published:
January 2013
Page Count:
90
Sponsoring Agency:
National Institute of Justice (NIJ) Washington, DC 20531 NCJRS Photocopy Services Rockville, MD 20849-6000
Grant Number:
2010-DN-BX-K171
Sale Source:
NCJRS Photocopy Services Box 6000 Rockville, MD 20849-6000 United States of America
Document:
PDF
Type:
Report (Grant Sponsored); Report (Study/Research)
Format:
Document (Online)
Language:
English
Country:
United States of America
Annotation:
This research succeeded in extending the “depth” of forensic mitochondrial DNA (mtDNA) analysis, i.e., the ability to detect minor variants arising from mutations, but present at very low levels.
Abstract:
The combination of an enhanced DNA extraction technique, whole genome amplification of the DNA extracts, multiplexed PCR amplification reactions around the mtDNA genome, and direct sequencing of the DNA samples on the Illumina® MiSeq™ instrument resulted in mtDNA sequence information from hair shafts that matched that found in blood and buccal extracts from the same donors. Further developmental research and validation based on this approach and the data obtained should result in a significant improvement over current forensic DNA typing procedures. In this effort, researchers evaluated two newly emerging methods of DNA sequence analysis in order to obtain massively parallel mtDNA sequence information (deep sequencing) from hair, buccal, and blood samples. The expanded information revealed that once this new technology is implemented into casework practice, interpretational change in forensic mtDNA analysis, reflecting the amounts of information that are produced, are necessary. Deep sequencing is a window into a level of variation that is currently under-appreciated in forensic casework. Researchers found that the general level of sequence heteroplasmy present in hair shaft samples, compared to blood and buccal samples, is heightened, but not to a level that would undermine the utility of mtDNA sequencing of hair shaft samples in a forensic context. Using the kit Nextera-XT™ from Illumina, Inc. enabled researchers to directly process any double-stranded DNA, including amplicons, for deep sequencing in a much simpler and cost-effective manner. 21 figures, 35 tables, and extensive references
Main Term(s):
Criminology
Index Term(s):
DNA Typing; Forensic sciences; Hair and fiber analysis; Investigative techniques; NIJ grant-related documents; Suspect identification
To cite this abstract, use the following link: http://www.ncjrs.gov/App/publications/abstract.aspx?ID=269378