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NCJ Number: 211280 Add to Shopping cart Find in a Library
Title: Alu-based, MGB Eclipse(TM) Real-Time PCR Method for Quantitation of Human DNA in Forensic Samples
Journal: Journal of Forensic Sciences  Volume:50  Issue:5  Dated:September 2005  Pages:1081-1090
Author(s): Janice A. Nicklas Ph.D.; Eric Buel Ph.D.
Date Published: September 2005
Page Count: 10
Sponsoring Agency: National Institute of Justice (NIJ)
Washington, DC 20531
Grant Number: 2000-IJ-CX-K012
Type: Test/Measurement
Format: Article
Language: English
Country: United States of America
Annotation: This paper describes the development of a real-time Alu sequence-based assay using MGB Eclipse primers and probes for the quantitation of human DNA in forensic samples.
Abstract: DNA isolated from crime scene samples must be quantitated to determine the amount of human DNA present. In attempting to develop faster, cheaper, and more quantitative methods, the forensic community has identified real-time PCR methods as having the advantages of being fast and quantitative with much less hands-on time than prior slot blot methods. This study describes the development and optimization of a MGB Eclipse assay, the Alu sequence-based assay, a real-time PCR method for the quantitation of human DNA. Most experiments were performed with a human DNA standard purchased from Promega in Wisconsin. DNA was isolated from a number of samples to evaluate the assay: blood spots from five DNA databank samples, five samples from control blood spotted on denim or colored cloth, control blood placed on five surfaces, a control blood spot placed in the dark for 3 months, two male and two female fractions from sexual assault cases, and two samples from a proficiency test. The primary advantage found for the MGB Eclipse real-time Alu assay was its much greater dynamic range. The Eclipse assay has the advantage of lower cost than the commercial kits and the greatest sample concentration range of published assays, additionally; it has simplicity, speed, less hands-on-time, and automated quantitation. Figures, tables, and references
Main Term(s): DNA fingerprinting
Index Term(s): Blood stains; Blood/body fluid analysis; Evidence identification; Forensic sciences; Suspect identification
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