Optimization of Human mtDNA Control Region Sequencing for Forensic Applications

• NCJ Number: 246022
• Journal: Journal of Forensic Sciences Volume: 59 Issue: 4 Dated: July 2014 Pages: 1057-1063
• Author(s): Veronique Bourdon Ph.D.; Carolyn Ng B.S.; Jessica Harris M.S.; Mechthild Prinz Ph.D.; Eli Shapiro Ph.D.
• Date Published: July 2014
• Length: 7 pages
• Publication Format: Article
• Publication Type: Report (Study/Research)
• Language: English
• Country: United States of America

Annotation

Sequencing mitochondrial DNA hypervariable regions I and II (HVI and HVII) is useful in forensic missing person and unidentified remains cases.

Abstract

Sequencing mitochondrial DNA hypervariable regions I and II (HVI and HVII) is useful in forensic missing person and unidentified remains cases. Improvements in ease and sensitivity of testing will yield results from more samples in a timely fashion. Routinely, amplification of HVI and HVII is followed by Sanger sequencing using the BigDye Terminator v3.1 Cycle Sequencing kit (Applied Biosystems) using 4 ìL of ready reaction mix (RRM). Each sequencing reaction is then purified through column filtration before capillary electrophoresis. Using lower amounts of RRM (2 ìL or 1 ìL) and purification using BigDye XTerminator (Applied Biosystems) instead of columns showed no loss of sequence length and increased the quality and the sensitivity of testing, allowing HVI and HVII typing from mitochondrial genome equivalent to 125 fg of nuclear DNA, or 100 pg of HVI/HVII amplicons. Using this methodology, testing can be completed in 1 day, and the cost of testing is reduced. Abstract published by arrangement with Wiley.