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Improved MtDNA Sequence Analysis of Forensic Remains Using a "Mini-Primer Set" Amplification Strategy

NCJ Number
187606
Journal
Journal of Forensic Sciences Volume: 46 Issue: 2 Dated: March 2001 Pages: 247-253
Author(s)
Matthew N. Gabriel M.F.S; Edwin F. Huffine M.S.; John H. Ryan Ph.D.; Mitchell M. Holland Ph.D.; Thomas J. Parsons Ph.D.
Date Published
March 2001
Length
7 pages
Annotation
This paper reports on an improved approach to forensic mitochondrial DNA (mtDNA) analysis of hyper-variable regions 1 and 2 (HV1/HV2) in highly degraded specimens.
Abstract
Mitochondrial DNA 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 shown 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, the research reported in this paper involved a "mini-primer set" (MPS) amplification strategy that consisted of four overlapping products that spanned each of the HV regions and ranged from 126 bp to 170 bp, with an average size of 141 bp. For this study, 11 extracts that represented a range of sample quality were prepared from nonprobative forensic specimens. The research showed a significant increase in MPS amplification success when compared to testing methods that use ~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. 2 tables, 3 figures, and 13 references