Simultaneous Detection of Human Mitochondrial DNA and Nuclear-Inserted Mitochondrial-Origin Sequences (NumtS) Using Forensic mtDNA Amplification Strategies and Pyrosequencing Technology

• NCJ Number: 246023
• Journal: Journal of Forensic Sciences Volume: 59 Issue: 4 Dated: July 2014 Pages: 1064-1073
• Author(s): Brittania J. Bintz M.S.; Groves B. Dixon M.S.; Mark R. Wilson Ph.D.
• Date Published: July 2014
• Length: 10 pages
• Publication Format: Article
• Publication Type: Report (Study/Research)
• Language: English
• Country: United States of America

Annotation

Next-generation sequencing technologies enable the identification of minor mitochondrial DNA variants with higher sensitivity than Sanger methods, allowing for enhanced identification of minor variants.

Abstract

Next-generation sequencing technologies enable the identification of minor mitochondrial DNA variants with higher sensitivity than Sanger methods, allowing for enhanced identification of minor variants. In this study, mixtures of human mtDNA control region amplicons were subjected to pyrosequencing to determine the detection threshold of the Roche GS Junior instrument (Roche Applied Science, Indianapolis, IN). In addition to expected variants, a set of reproducible variants was consistently found in reads from one particular amplicon. A BLASTn search of the variant sequence revealed identity to a segment of a 611-bp nuclear insertion of the mitochondrial control region (NumtS) spanning the primer-binding sites of this amplicon (Nature 1995;378:489). Primers (Hum Genet 2012;131:757; Hum Biol 1996;68:847) flanking the insertion were used to confirm the presence or absence of the NumtS in buccal DNA extracts from twenty donors. These results further our understanding of human mtDNA variation and are expected to have a positive impact on the interpretation of mtDNA profiles using deep-sequencing methods in casework. Abstract published by arrangement with Wiley.