Barbaro, A.; Falcone, G.; La Marca, A.; Barbaro, A. Use of the Investigator ESSplex SE QS Kit (QIAGEN) at Half PCR Reaction Volumes for the Analysis of Forensic Samples. Forensic Sci.2024, 4, 152-163.
Barbaro, A.; Falcone, G.; La Marca, A.; Barbaro, A. Use of the Investigator ESSplex SE QS Kit (QIAGEN) at Half PCR Reaction Volumes for the Analysis of Forensic Samples. Forensic Sci. 2024, 4, 152-163.
Barbaro, A.; Falcone, G.; La Marca, A.; Barbaro, A. Use of the Investigator ESSplex SE QS Kit (QIAGEN) at Half PCR Reaction Volumes for the Analysis of Forensic Samples. Forensic Sci.2024, 4, 152-163.
Barbaro, A.; Falcone, G.; La Marca, A.; Barbaro, A. Use of the Investigator ESSplex SE QS Kit (QIAGEN) at Half PCR Reaction Volumes for the Analysis of Forensic Samples. Forensic Sci. 2024, 4, 152-163.
Abstract
Background: the Investigator ESSplex SE QS Kit (Qiagen) is a next generation PCR kit that, in 60 minutes, amplifies 17 STRs markers, including the 5 European Standard Set (ESS) loci, the SE33 marker and the locus Amelogenin for sex determination. Two Quality Sensors (QS1 and QS2) are co-amplified to check PCR performance. Since forensic laboratories carry out annually hundreds of DNA typing, we verified the kit performance using half reaction volumes with the aim to improve the number of samples that may amplified with a kit, reducing consequently the laboratory costs.
Methods: in the study, that is intended as a technical note and not an internal validation, were analyzed control samples (oral swabs) with known DNA profiles and 40 real caseworks samples.
Results: we observed that reducing the total reaction volume, while keeping unaltered all components ratio’s, yields DNA profiles comparable to the ones obtained using standard reaction volumes and with alleles peaks higher than with standard volumes..
Conclusions: the use of half volumes for PCR amplification allows the analysis of a wider number of samples than the standard protocol, reducing laboratory costs while maintaining unaltered the quality level of the analysis.
Keywords
ESSplexQS; STR multiplex; half reaction volumes
Subject
Biology and Life Sciences, Biology and Biotechnology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
