Quantification of Human Mitochondrial DNA Using Synthesized DNA Standards

• NCJ Number: 236874
• Journal: Journal of Forensic Sciences Volume: 56 Issue: 6 Dated: November 2011 Pages: 1457;-1463;
• Author(s): Mark F. Kavlick, B.S.; Helen S. Lawrence, M.S.; R. Travis Merritt, B.S.; Constance Fisher, Ph.D.; Alice Isenberg, Ph.D.; James M. Robertson, Ph.D.; Bruce Budowle, Ph.D.
• Date Published: November 2011
• Length: 7 pages
• Corporate Author: US Dept of Justice Federal Bureau of Investigation
• Publication Format: Article
• Publication Type: Report (Study/Research)
• Language: English
• Country: United States of America

Annotation

This study examined real-time quantitative PCR assay developed to assess mitochondrial DNA (mtDNA), which utilized a duplex, synthetic DNA to ensure optimal quality assurance and quality control.

Abstract

Successful mitochondrial DNA (mtDNA) forensic analysis depends on sufficient quantity and quality of mtDNA. A real-time quantitative PCR assay was developed to assess such characteristics in a DNA sample, which utilizes a duplex, synthetic DNA to ensure optimal quality assurance and quality control. The assay's 105-base pair target sequence facilitates amplification of degraded DNA and is minimally homologous to nonhuman mtDNA. The primers and probe hybridize to a region that has relatively few sequence polymorphisms. The assay can also identify the presence of PCR inhibitors and thus indicate the need for sample repurification. The results show that the assay provides information down to 10 copies and provides a dynamic range spanning seven orders of magnitude. Additional experiments demonstrated that as few as 300 mtDNA copies resulted in successful hypervariable region amplification, information that permits sample conservation and optimized downstream PCR testing. The assay described is rapid, reliable, and robust. (Published Abstract)