1. The digital MIQE guidelines update : minimum information for publication of quantitative digital PCR experiments for 2020Jim F. Huggett, Alexandra S. Whale, Ward De Spiegelaere, Afif M. Abdel Nour, Young-Kyung Bae, Vladimír Beneš, Dan Burke, Megan Cleveland, Philippe Corbisier, Alison S. Devonshire, Lianhua Dong, Daniela Drandi, Carole A. Foy, Jeremy A. Garson, Hua-Jun He, Jan Hellemans, Mikael Kubista, Antoon Lievens, Mike G. Makrigiorgos, Mojca Milavec, Reinhold D. Mueller, Tania Nolan, Denise M. O'Sullivan, Michael W. Pfaffl, Stefan Rödiger, Erica L. Romsos, Gregory L. Shipley, Valérie Taly, Andreas Untergasser, Carl T. Wittwer, Stephen A. Bustin, Jo Vandesompele, 2020, review article Abstract: Digital PCR (dPCR) has developed considerably since the publication of the Minimum Information for Publication of Digital PCR Experiments (dMIQE) guidelines in 2013, with advances in instrumentation, software, applications, and our understanding of its technological potential. Yet these developments also have associated challenges; data analysis steps, including threshold setting, can be difficult and preanalytical steps required to purify, concentrate, and modify nucleic acids can lead to measurement error. To assist independent corroboration of conclusions, comprehensive disclosure of all relevant experimental details is required. To support the community and reflect the growing use of dPCR, we present an update to dMIQE, dMIQE2020, including a simplified dMIQE table format to assist researchers in providing key experimental information and understanding of the associated experimental process. Adoption of dMIQE2020 by the scientific community will assist in standardizing experimental protocols, maximize efficient utilization of resources, and further enhance the impact of this powerful technology.
Published in DiRROS: 06.08.2024; Views: 27; Downloads: 77
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2. Digital PCR for the characterization of reference materialsMegan H. Cleveland, Hua-Jun He, Mojca Milavec, Young-Kyung Bae, Peter M. Vallone, Jim F. Huggett, 2024, review article Abstract: Well-characterized reference materials support harmonization and accuracy when conducting nucleic acid-based tests (such as qPCR); digital PCR (dPCR) can measure the absolute concentration of a specific nucleic acid sequence in a background of non-target sequences, making it ideal for the characterization of nucleic acid-based reference materials. National Metrology Institutes are increasingly using dPCR to characterize and certify their reference materials, as it offers several advantages over indirect methods, such as UV-spectroscopy. While dPCR is gaining widespread adoption, it requires optimization and has certain limitations and considerations that users should be aware of when characterizing reference materials. This review highlights the technical considerations of dPCR, as well as its role when developing and characterizing nucleic acid-based reference materials.
Keywords: digital PCR, dPCR, reference materials, UV-spectroscopy
Published in DiRROS: 03.06.2024; Views: 271; Downloads: 103
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