Quantification of human mitochondrial DNA Using synthesized DNA Standards

Mark F. Kavlick, Helen S. Lawrence, R. Travis Merritt, Constance Fisher, Alice Isenberg, James M. Robertson, Bruce Budowle

Research output: Contribution to journalArticle

35 Scopus citations

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. 2011 American Academy of Forensic Sciences. Published 2011. This article is a U.S. Government work and is in the public domain in the U.S.A..

Original languageEnglish
Pages (from-to)1457-1463
Number of pages7
JournalJournal of Forensic Sciences
Volume56
Issue number6
DOIs
StatePublished - 1 Nov 2011

Keywords

  • Assay
  • DNA quality
  • DNA quantification
  • Forensic science
  • Mitochondrial DNA
  • Quantitative PCR
  • Synthetic DNA standards

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    Kavlick, M. F., Lawrence, H. S., Merritt, R. T., Fisher, C., Isenberg, A., Robertson, J. M., & Budowle, B. (2011). Quantification of human mitochondrial DNA Using synthesized DNA Standards. Journal of Forensic Sciences, 56(6), 1457-1463. https://doi.org/10.1111/j.1556-4029.2011.01871.x