Skip to main content
U.S. flag

An official website of the United States government

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.

Demonstration of Rapid Multiplex PCR Amplification Involving 16 Genetic Loci

Published

Author(s)

Peter Vallone, Carolyn R. Steffen, John M. Butler

Abstract

Current forensic DNA typing is conducted in approximately eight to ten hours with steps including DNA extraction, quantitation, polymerase chain reaction (PCR) amplification of multiple short tandem repeat (STR) loci, capillary electrophoresis separation with fluorescence detection and data analysis, and DNA profile interpretation. The PCR amplification portion of the workflow typically takes approximately three hours with standard thermal cycling protocols.  Here we demonstrate a rapid cycling protocol that amplifies 15 STR loci and the sex-typing marker amelogenin from either Identifiler or PowerPlex 16 STR typing kits in less than 36 minutes.  This rapid protocol employs commercially available polymerases and the widely used GeneAmp 9700 thermal cycler. We observed complete concordance of STR allele calls between the rapid and standard thermal cycling protocols although there was incomplete adenylation at several of the loci examined and some PCR artifacts were detected. Using less than 1 ng of template DNA and 28 cycles, STR peaks for all loci were above a 100 relative fluorescent unit (RFU) detection threshold with fully adequate inter-locus balance and heterozygote peak height ratios of greater than 0.80.
Citation
Forensic Science International
Volume
3
Issue
1

Keywords

rapid PCR, PCR, STR, DNA typing, multiplex PCR, PowerPlex 16, Identifiler

Citation

Vallone, P. , Steffen, C. and Butler, J. (2008), Demonstration of Rapid Multiplex PCR Amplification Involving 16 Genetic Loci, Forensic Science International, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=832219 (Accessed January 26, 2022)
Created November 30, 2008, Updated February 19, 2017