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NCJRS Abstract

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NCJ Number: 228279 Add to Shopping cart Find in a Library
Title: Development of a Multiplex PCR and Linear Array Probe Assay Targeting Informative Polymorphisms Within the Entire Mitochondrial Genome
Author(s): Cassandra D. Calloway; Sarah M. Stuart; Henry A. Erlich
Date Published: 2009
Page Count: 115
Sponsoring Agency: National Institute of Justice (NIJ)
Washington, DC 20531
National Institute of Justice/NCJRS
Rockville, MD 20849
NCJRS Photocopy Services
Rockville, MD 20849-6000
Grant Number: 2005-MU-BX-K074
Sale Source: National Institute of Justice/NCJRS
Box 6000
Rockville, MD 20849
United States of America

NCJRS Photocopy Services
Box 6000
Rockville, MD 20849-6000
United States of America
Document: PDF
Type: Report (Study/Research)
Format: Document
Language: English
Country: United States of America
Annotation: In order to increase the information provided by mitochondrial DNA (mtDNA) analysis by targeting additional sequence polymorphisms outside the HVI/II regions, the project described in this report developed a highly sensitive, easy-to-use, 5-plex and 10-plex PCR and linear array assay for simultaneous analysis of polymorphic regions in the noncoding and coding regions.
Abstract: This assay targets 61 polymorphic sites distributed throughout the mitochondrial genome, using 15 primer pairs and 105 sequence-specific oligonucleotide probes immobilized in lines on a nylon membrane. The 5-plex PCR is used to amplify 5 regions that range in size from 314-444 bp; the 10-plex PCR is used to amplify 10 regions that range in size from 103-183 bp. The 5-plex probe panel consists of 59 probes and targets variation at 14 HVI sites, 11 HVII sites, 8 coding region (CR) sites, and 2 variable region I (VRI) sites. The 10-plex probe panel consists of 46 probes and targets 17 CR sites, 4 VRI sites, and five VRII sites. A population study was conducted in order to determine the power of discrimination for the new expanded HV+ array. In order to determine the assay limitations, a developmental validation study was conducted. A sensitivity study showed that both the 5-plex and 10-plex assays were highly sensitive assays, with the 10-plex being more sensitive. Additional developmental studies - including reproducibility, precision and accuracy, and environmental samples - were also completed. In addition, a procedure was validated for automating the typing assay, and scanning and interpretation software were modified for use with the 5-plex and 10-plex assays. Up to 48 samples can be typed manually or automated in less than 2 hours. The data can now be quickly analyzed with the Strip Scan Mitotyper software. 19 figures, 6 tables, and 42 references
Main Term(s): Criminology
Index Term(s): DNA fingerprinting; Forensic sciences; Investigative techniques; Suspect identification; Victim identification
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