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Volume 23
Issue 15
10.3390/ijms23158486
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Technical Note

Maximising the Use of Scarce qPCR Master Mixes

by
Stephen Bustin
1,*,
Claire Bustin
2,
Sara Kirvell
1,
Tania Nolan
1,
Reinhold Mueller
3 and
Gregory Shipley
4
1
Medical Technology Research Centre, Anglia Ruskin University, Chelmsford CM1 1SQ, UK
2
Royal Wolverhampton NHS Trust, Wolverhampton WV10 0QP, UK
3
RM Consulting, San Diego, CA 92037, USA
4
Shipley Consulting, Vancouver, WA 98682, USA
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2022, 23(15), 8486; https://doi.org/10.3390/ijms23158486
Submission received: 22 June 2022 / Revised: 24 July 2022 / Accepted: 28 July 2022 / Published: 30 July 2022
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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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.
Keywords: PCR; qPCR; sensitivity; PCR efficiency; molecular diagnostics PCR; qPCR; sensitivity; PCR efficiency; molecular diagnostics

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MDPI and ACS Style

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, 23, 8486. https://doi.org/10.3390/ijms23158486

AMA Style

Bustin S, Bustin C, Kirvell S, Nolan T, Mueller R, Shipley G. Maximising the Use of Scarce qPCR Master Mixes. International Journal of Molecular Sciences. 2022; 23(15):8486. https://doi.org/10.3390/ijms23158486

Chicago/Turabian Style

Bustin, Stephen, Claire Bustin, Sara Kirvell, Tania Nolan, Reinhold Mueller, and Gregory Shipley. 2022. "Maximising the Use of Scarce qPCR Master Mixes" International Journal of Molecular Sciences 23, no. 15: 8486. https://doi.org/10.3390/ijms23158486

APA Style

Bustin, S., Bustin, C., Kirvell, S., Nolan, T., Mueller, R., & Shipley, G. (2022). Maximising the Use of Scarce qPCR Master Mixes. International Journal of Molecular Sciences, 23(15), 8486. https://doi.org/10.3390/ijms23158486

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