Highlights
- •An efficient enzyme is crucial in forensic DNA analysis if inhibitors are present.
- •KAPA3G Plant and KAPA2G Robust enzymes have high resistance to certain inhibitors.
- •The KAPA polymerases counteract inhibitors in analysis of challenging samples.
Abstract
Inhibitors of polymerase chain reaction (PCR) amplification often present a challenge
in forensic investigations of e.g., terrorism, missing persons, sexual assaults and
other criminal cases. Such inhibitors may be counteracted by dilution of the DNA extract,
using different additives, and selecting an inhibitory resistant DNA polymerase. Additionally,
DNA in forensic samples is often present in limited amounts and degraded, requiring
special analyses of short nuclear targets or mitochondrial DNA. The present study
evaluated the enzymes AmpliTaq Gold, HotStarTaq Plus, KAPA3G Plant, and KAPA2G Robust,
with regard to their ability to overcome inhibitory effects. Our data showed that
diluting the extracts and adding bovine serum albumin may increase the yield of the
PCR product. However, the largest impact was observed when alternative enzymes were
utilized, instead of the commonly used AmpliTaq Gold. KAPA2G Robust presented the
highest amplification efficiency in the presence of the inhibitor ammonium nitrate.
Moreover, the KAPA3G Plant enzyme had the highest efficiency in amplifying degraded
DNA from old buried bone material. KAPA3G Plant and KAPA2G Robust may thus be useful
for counteracting inhibitors and improving the analysis of challenging samples.
Keywords
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Article info
Publication history
Published online: June 01, 2016
Accepted:
May 29,
2016
Received in revised form:
April 25,
2016
Received:
February 23,
2016
Identification
Copyright
© 2016 Elsevier Ireland Ltd. All rights reserved.
