Journal
FORENSIC SCIENCE INTERNATIONAL GENETICS SUPPLEMENT SERIES
Volume 5, Issue -, Pages E5-E6Publisher
ELSEVIER IRELAND LTD
DOI: 10.1016/j.fsigss.2015.09.003
Keywords
Dithiothreitol; Fluorescence quenching; Passive reference dye; PCR inhibition; qPCR
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Forensic DNA analysis is partly limited by PCR-inhibitory compounds present in the DNA extracts. Generally, these inhibitors disturb amplification, i.e., the production of amplicons. We have found that dithiothreitol (DTT) from the DNA extraction process can cause another type of real-time PCR disturbance, i.e., inhibition of signal detection through fluorescence quenching. DNA extracts containing DTT substantially quenched the passive reference signal in the Quantifiler HP DNA Quantification kit. This quenching resulted in overestimation of DNA concentrations, as target DNA signals are normalized to the passive reference signal. (C) 2015 Elsevier Ireland Ltd. All rights reserved.
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