Maximising the Use of Scarce qPCR Master Mixes

Bustin, Stephen ORCID: https://orcid.org/0000-0003-1870-6098, Bustin, Claire, Kirvell, Sara, Nolan, Tania, Mueller, Reinhold D. and Shipley, Gregory L. (2022) Maximising the Use of Scarce qPCR Master Mixes. International Journal of Molecular Sciences, 23. p. 8486. ISSN 1422-0067

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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.

Item Type:	Journal Article
Keywords:	PCR, qPCR, sensitivity, PCR efficiency, Molecular diagnostics
Faculty:	COVID-19 Research Collection Faculty of Health, Education, Medicine & Social Care
Depositing User:	Dr Catherine Pearson
Date Deposited:	16 Sep 2022 08:57
Last Modified:	16 Sep 2022 08:57
URI:	https://arro.anglia.ac.uk/id/eprint/707924

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