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Maximising the Use of Scarce qPCR Master Mixes
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 chemic...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9368830/ https://www.ncbi.nlm.nih.gov/pubmed/35955620 http://dx.doi.org/10.3390/ijms23158486 |
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author | Bustin, Stephen Bustin, Claire Kirvell, Sara Nolan, Tania Mueller, Reinhold Shipley, Gregory |
author_facet | Bustin, Stephen Bustin, Claire Kirvell, Sara Nolan, Tania Mueller, Reinhold Shipley, Gregory |
author_sort | Bustin, Stephen |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9368830 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-93688302022-08-12 Maximising the Use of Scarce qPCR Master Mixes Bustin, Stephen Bustin, Claire Kirvell, Sara Nolan, Tania Mueller, Reinhold Shipley, Gregory Int J Mol Sci Technical Note 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. MDPI 2022-07-30 /pmc/articles/PMC9368830/ /pubmed/35955620 http://dx.doi.org/10.3390/ijms23158486 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Technical Note Bustin, Stephen Bustin, Claire Kirvell, Sara Nolan, Tania Mueller, Reinhold Shipley, Gregory Maximising the Use of Scarce qPCR Master Mixes |
title | Maximising the Use of Scarce qPCR Master Mixes |
title_full | Maximising the Use of Scarce qPCR Master Mixes |
title_fullStr | Maximising the Use of Scarce qPCR Master Mixes |
title_full_unstemmed | Maximising the Use of Scarce qPCR Master Mixes |
title_short | Maximising the Use of Scarce qPCR Master Mixes |
title_sort | maximising the use of scarce qpcr master mixes |
topic | Technical Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9368830/ https://www.ncbi.nlm.nih.gov/pubmed/35955620 http://dx.doi.org/10.3390/ijms23158486 |
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