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NCJ Number: NCJ 210302   Add to Shopping cart   Find in a Library
Title: Quantitation of DNA for Forensic DNA Typing by qPCR (quantitative PCR): Singleplex and Multiple Modes for Nuclear and Mitochondrial Genomes, and the Y Chromosome
Author(s): Mark D. Timken ; Katie L. Swango ; Cristian Orrego ; Mavis Date Chong ; Martin R. Buoncristiani
Date Published: 2005
Page Count: 89
Sponsoring Agency: National Institute of Justice
US Department of Justice
Office of Justice Programs
United States of America
Grant Number: 2002-IJ-CX-K008
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: Research (Applied/Empirical)
Language: English
Country: United States of America
Annotation: This report presents three quantitation assays developed to improve the accuracy and precision of DNA quantification, as well as tools to increase the effectiveness of the analysis of complex forensic samples.
Abstract: The quantitation of DNA plays a central role in all areas and applications of forensic DNA analysis. The careful evaluation of the quantity of DNA extracted from biological samples is an imperative for DNA typing via PCR. However, the quantitation methods commonly employed in forensics are often less accurate, time consuming, and require the consumption of a substantial percentage of what may be a precious sample. In this report, supported by the U.S. Department of Justice, National Institute of Justice, the use of real-time quantitative PCR (qPCR), a method commonly employed for a variety of molecular diagnostic and biotechnology procedures, was proposed for the quantification of DNA in forensic samples. The intent is to develop qPCR assays to quantify the human nuclear genome, the human mitochondrial genome, and the human Y-chromosome, and to multiplex these assays in order to quantify more than one genome (or chromosome) in a single reaction. Three quantitation assays were developed based on real-time quantitative PCR (qPCR) methods and include: (1) a duplex qPCR assay that quantifies simultaneously the human nuclear and mitochondrial genomes; (2) a triplex assay for assessing the quantity and quality of human nuclear DNA; and (3) several singleplex assays to quantify human male (Y-chromosomal) DNA. All assays (tests) were developed utilizing 5’-hydrolysis detection chemistries for data collection on the ABI 7000 instrument. References
Main Term(s): DNA fingerprinting
Index Term(s): Testing and measurement ; Trace evidence ; Evidence identification and analysis ; Evaluation measures ; Effectiveness ; Forensics/Forensic Sciences ; NIJ grant-related documents
   
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https://www.ncjrs.gov/App/Publications/abstract.aspx?ID=210302

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