U.S. flag

An official website of the United States government, Department of Justice.

NCJRS Virtual Library

The Virtual Library houses over 235,000 criminal justice resources, including all known OJP works.
Click here to search the NCJRS Virtual Library

TWGDAM Validation of AmpFlSTR™ PCR Amplification Kits for Forensic DNA Casework

NCJ Number
193726
Journal
Journal of Forensic Sciences Volume: 47 Issue: 1 Dated: January 2002 Pages: 66-96
Author(s)
Cydne Holt Ph.D.; Martin Buoncristiani M.P.H; Jeanette M. Wallin M.P.H; Theresa Nguyen B.S.; Katherine D. Lazaruk Ph.D.; P. S. Walsh M.P.H
Date Published
2002
Length
31 pages
Annotation
This study examined the Technical Working Group on DNA Analysis Methods (TWGDAM) validation of AmpFlSTR™PCR Amplification Kits for forensic DNA casework.
Abstract
Short tandem repeat (STR) analysis were subjected to various scenarios in laboratory procedures that assessed reliability and identified potential limitations. These validation studies were designed as recommended by the TWGDAM and the DNA Advisory Board (DAB). Various DNA samples were amplified by the polymerase chain reaction (PCR) using AmpFlSTR™, Profiler Plus™, and COfiler™, detected with ABI Prism instrumentation and analyzed using GeneScan and Genotyper software. Data acquired in these studies reinforced an existing body of knowledge and expertise regarding application and interpretation of STR typing in the forensic science community. Consistent STR genotypes were detected in various body tissues and fluids. Inter-laboratory comparisons produced similar genotype results. Quantitative interpretational aids for DNA mixtures were characterized. Ability of the typing system to type potentially compromised samples reliably was evaluated. Nonprobative case evidentiary DNA was successfully amplified, genotyped, and interpreted. Potential limitations or cautionary factors in the interpretation of minimal fluorescence intensity were demonstrated. Differential amplification between loci was observed when PCR was inhibited; preferential amplification typically was not. Single AmpFlSTR locus amplification did not offer significant benefit over AmpFlSTR multiplexing, even in cases of DNA degradation or PCR inhibition. During rigorous evaluation, AmpFlSTR PCR Amplification Kits reproducibly yielded sensitive and locus-specific results, as called for in routine forensic analyses. 34 References