Abstract
Current forensic DNA typing is conducted in approximately 8–10 h. Steps include DNA extraction, quantification, polymerase chain reaction (PCR) amplification
of multiple short tandem repeat (STR) loci, capillary electrophoresis separation with
fluorescence detection, data analysis and DNA profile interpretation. The PCR amplification
portion of the workflow typically takes approximately 3 h with standard thermal cycling protocols. Here we demonstrate a rapid cycling protocol
that amplifies 15 STR loci and the sex-typing marker amelogenin from the Identifiler
STR typing kit in less than 36 min. This rapid protocol employs commercially available polymerases and the widely
used GeneAmp 9700 thermal cycler. Complete concordance of STR allele calls (for 60
samples) between the rapid and standard thermal cycling protocols were observed although
there was incomplete adenylation at several of the loci examined and some PCR artifacts
were detected. Using less than 750 pg of template DNA and 28 cycles, STR peaks for all loci were above a 150 relative
fluorescent unit (RFU) detection threshold with fully adequate inter-locus balance
and heterozygote peak height ratios of greater than 0.84.
Keywords
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Article info
Publication history
Published online: October 27, 2008
Accepted:
September 14,
2008
Received in revised form:
September 10,
2008
Received:
August 4,
2008
Footnotes
☆Certain commercial equipment, instruments and materials are identified in order to specify experimental procedures as completely as possible. In no case does such identification imply a recommendation or endorsement by the National Institute of Standards and Technology nor does it imply that any of the materials, instruments or equipment identified are necessarily the best available for the purpose.
Identification
Copyright
Published by Elsevier Inc.
