Maximising the use of scarce qPCR master mixes.
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Authors
Bustin, Claire
Issue Date
2022-07-30
Journal
Type
Peer-Reviewed Publication
Keywords
COVID-19
Polymerase chain reaction
Sensitivity
Specificity
PCR tests
Molecular diagnostics
Polymerase chain reaction
Sensitivity
Specificity
PCR tests
Molecular diagnostics
Journal Title
International Journal of Molecular Sciences
Volume
23
Issue
15
Begin page
8486
End page
Abstract
The COVID-19 pandemic resulted in a universal, immediate, and vast demand for comprehensive molecular diagnostic testing, especially real-time quantitative (qPCR)-based methods. This rapidly triggered a global shortage of testing capacity, equipment, and reagents. Even today, supply times for chemicals from date of order to delivery are often much longer than pre-pandemic. Furthermore, many companies have ratcheted up the price for minimum volumes of reaction master mixes essential for qPCR assays, causing additional problems for academic laboratories often operating on a shoestring. We have validated two strategies that stretch reagent supplies and, whilst particularly applicable in case of scarcity, can readily be incorporated into standard qPCR protocols, with appropriate validation. The first strategy demonstrates equivalent performance of a selection of "past expiry date" and newly purchased master mixes. This approach is valid for both standard and fast qPCR protocols. The second validates the use of these master mixes at less than 1x final concentration without loss of qPCR efficiency or sensitivity.
Citation
Bustin S, Bustin C, Kirvell S, Nolan T, Mueller R, Shipley G. Maximising the Use of Scarce qPCR Master Mixes. Int J Mol Sci. 2022 Jul 30;23(15):8486. doi: 10.3390/ijms23158486. PMID: 35955620; PMCID: PMC9368830.