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NCJRS Abstract

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  NCJ Number: NCJ 226638   Add to Shopping cart   Find in a Library
  Title: Mitochondrial DNA Analysis by Denaturing Liquid Chromatography for the Separation of Mixtures in Forensic Samples
  Document URL: PDF 
  Author(s): Phillip B. Danielson
  Date Published: 2009
  Page Count: 106
  Annotation: This report describes the methodology and presents the results of research that involved the developmental validation of denaturing high-performance liquid chromatography (DHPLC) for the rapid and accurate resolution of mitochondrial DNA (mtDNA) mixtures.
  Abstract: The research shows that DHPLC is an accurate method for rapid sequence-specific fractionation of heteroplasmic or multicontributor DNA mixtures prior to sequencing. Fractionation allows the mtDNA sequence of individual contributors to be unambiguously determined without secondary amplification or excessive manipulation. The demonstrated reproducibility of electrophoretic sequence profiles and correlation with changes in DNA quantity ratios makes it possible to determine the linkage phase of individual amplicons from even partially resolved mtDNA mixtures. In addition, DHPLC can be used to streamline the entire mtDNA analysis process flow by enabling analysts to assess sample complexity, accurately quantify PCR product yield without gels, purify target amplicons to facilitate improved dye terminator labeling, and confirm negative controls without sequencing. This approach has also been validated by using casework-type samples. Detailed descriptions of methods and materials address mtDNA extraction and sequencing and DHPLC analysis. 31 figures, 8 tables, 37 references, and appended draft standard operating procedures for DHPLC analysis of mtDNA
  Main Term(s): Criminology
  Index Term(s): Chromatography ; Victim identification ; Suspect identification ; Forensics/Forensic Sciences ; Investigative techniques ; DNA fingerprinting ; NIJ final report
  Sponsoring Agency: National Institute of Justice (NIJ)
US Department of Justice
Office of Justice Programs
United States of America
  Grant Number: 2003-IJ-CX-K104
  Sale Source: National Institute of Justice/NCJRS
Box 6000
Rockville, MD 20849
United States of America

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  Type: Report (Study/Research)
  Country: United States of America
  Language: English
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