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Maximising the Use of Scarce qPCR Master Mixes

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journal contribution
posted on 2023-07-26, 15:59 authored by Stephen Bustin, Claire Bustin, Sara Kirvell, Tania Nolan, Reinhold D. Mueller, Gregory L. Shipley
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.

History

Refereed

  • Yes

Volume

23

Page range

8486

Publication title

International Journal of Molecular Sciences

ISSN

1422-0067

Publisher

MDPI

File version

  • Published version

Language

  • eng

Legacy posted date

2022-09-16

Legacy creation date

2022-09-16

Legacy Faculty/School/Department

COVID-19 Research Collection

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