Improved mtDNA sequence analysis of forensic remains using a "mini-primer set" amplification strategy

M. N. Gabriel, E. F. Huffine, J. H. Ryan, Mitchell Mark Holland, Thomas John Parsons

Research output: Contribution to journalArticle

100 Citations (Scopus)

Abstract

Mitochondrial DNA (mtDNA) analysis of highly degraded skeletal remains is often used for forensic identification due largely to the high genome copy number per cell. Literature from the "ancient DNA" field has shown that highly degraded samples contain populations of intact DNA molecules that are severely restricted in size (1-4). Hand et al, have demonstrated the targeting and preferential amplification of authentic human DNA sequences with small amplicon products of 150 bp or less (1,2). Given this understanding of ancient DNA preservation and amplification, we report an improved approach to forensic mtDNA analysis of hyper-variable regions I and 2 (HV1/HV2) in highly degraded specimens. This "mini-primer set" (MPS) amplification strategy consists of four overlapping products that span each of the HV regions and range from 126 to 170 bp, with an average size of 141 bp. For this study, 11 extracts representing a range of sample quality were prepared from nonprobative forensic specimens. We demonstrate a significant increase in MPS amplification success when compared to testing methods using ∼250 bp amplicons. Further, 16 of 17 independent amplifications previously "unreported" due to mixed sequences provided potentially reportable sequence data from a single, authentic template with MPS testing.

Original languageEnglish (US)
Pages (from-to)247-253
Number of pages7
JournalJournal of Forensic Sciences
Volume46
Issue number2
StatePublished - Apr 28 2001

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genetic test
Mitochondrial DNA
DNA Sequence Analysis
Cell Count
Genome
DNA
Population
Ancient DNA
Body Remains
literature

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Genetics

Cite this

Gabriel, M. N. ; Huffine, E. F. ; Ryan, J. H. ; Holland, Mitchell Mark ; Parsons, Thomas John. / Improved mtDNA sequence analysis of forensic remains using a "mini-primer set" amplification strategy. In: Journal of Forensic Sciences. 2001 ; Vol. 46, No. 2. pp. 247-253.
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Improved mtDNA sequence analysis of forensic remains using a "mini-primer set" amplification strategy. / Gabriel, M. N.; Huffine, E. F.; Ryan, J. H.; Holland, Mitchell Mark; Parsons, Thomas John.

In: Journal of Forensic Sciences, Vol. 46, No. 2, 28.04.2001, p. 247-253.

Research output: Contribution to journalArticle

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