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Open Access 01-01-2021 | Original Article

Impact of metal ions on PCR inhibition and RT-PCR efficiency

Authors: Agnieszka Kuffel, Alexander Gray, Niamh Nic Daeid

Published in: International Journal of Legal Medicine | Issue 1/2021

Abstract

Inhibition of PCR by metal ions can pose a serious challenge in the process of forensic DNA analysis. Samples contaminated with various types of metal ions encountered at crime scenes include swabs from metal surfaces such as bullets, cartridge casings, weapons (including guns and knives), metal wires and surfaces as well as bone samples which contain calcium. The mechanism behind the impact of metal ions on DNA recovery, extraction and subsequent amplification is not fully understood. In this study, we assessed the inhibitory effects of commonly encountered metals on DNA amplification. Of the nine tested metals, zinc, tin, iron(II) and copper were shown to have the strongest inhibitory properties having IC50 values significantly below 1 mM. In the second part of the study, three commercially available DNA polymerases were tested for their susceptibility to metal inhibition. We found that KOD polymerase was the most resistant to metal inhibition when compared with Q5 and Taq polymerase. We also demonstrate how the calcium chelator ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) can be used as an easy and non-destructive method of reversing calcium-induced inhibition of PCR reactions.

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Title: Impact of metal ions on PCR inhibition and RT-PCR efficiency
Authors: Agnieszka Kuffel
Alexander Gray
Niamh Nic Daeid
Publication date: 01-01-2021
Publisher: Springer Berlin Heidelberg
Published in: International Journal of Legal Medicine / Issue 1/2021
Print ISSN: 0937-9827
Electronic ISSN: 1437-1596
DOI: https://doi.org/10.1007/s00414-020-02363-4

At a glance: The STEP trials

Springer Medicine

Impact of metal ions on PCR inhibition and RT-PCR efficiency

Abstract

At a glance: The STEP trials

Springer Medicine

Abstract

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