Bustin_et_al_2022.pdf (4.33 MB)
Maximising the Use of Scarce qPCR Master Mixes
journal contribution
posted on 2023-07-26, 15:59 authored by Stephen Bustin, Claire Bustin, Sara Kirvell, Tania Nolan, Reinhold D. Mueller, Gregory L. ShipleyThe 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.
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Refereed
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23Page range
8486Publication title
International Journal of Molecular SciencesISSN
1422-0067External DOI
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MDPIFile version
- Published version
Language
- eng
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2022-09-16Legacy creation date
2022-09-16Legacy Faculty/School/Department
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