Abstract
The analysis of short tandem repeats is one of the most powerful tools in forensic
genetics. Forensic practice sometimes requires the individualization of samples that
may contain only highly degraded nuclear DNA, mitochondrial DNA or PCR inhibitors
that hamper DNA amplification. We designed a new multiplex PCR with reduced size amplicons
(<200 bp), providing a double sex determination (amelogenin plus two Y-STRs), the detection
of two autosomal markers and the amplification of mitochondrial specific fragments
from the hypervariable region I (HVI). Additionally, a quality sensor was developed
to check for the presence of any PCR inhibitors.
The new multiplex PCR shows a reproducible detection threshold down to 25 pg and gives signals even out of highly degraded materials. All signals are reproducible
and reliable as it could be shown in comparison to results from commercially available
STR multiplex-PCRs. In no case DNA fragments were detectable using any other assay
when the quality sensor was not detectable.
There was a good correlation between detection of mitochondrial specific fragments
in the multiplex-PCR and success of subsequent sequencing of HVI region. The same
could be shown for STR analysis: Most samples successfully analyzed in our PCR yielded
at least a partial STR profile using a commercial STR kit.
We present an assay that allows an easy, reliable, and cost efficient evaluation of
DNA sample quality combined with a first rough sample individualization and sex determination
suitable for forensic purposes. This assay may help the forensic lab personnel to
decide on further sample processing.
Keywords
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Article info
Publication history
Published online: January 09, 2009
Accepted:
November 14,
2008
Received in revised form:
October 15,
2008
Received:
March 18,
2008
Identification
Copyright
© 2008 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
