Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

FlashPCR: Revolutionising qPCR by Accelerating Amplification through Low ∆T Protocols

Version 1 : Received: 17 January 2024 / Approved: 18 January 2024 / Online: 18 January 2024 (12:01:04 CET)

A peer-reviewed article of this Preprint also exists.

Bustin, S.A.; Kirvell, S.; Nolan, T.; Shipley, G.L. FlashPCR: Revolutionising qPCR by Accelerating Amplification through Low ∆T Protocols. Int. J. Mol. Sci. 2024, 25, 2773. Bustin, S.A.; Kirvell, S.; Nolan, T.; Shipley, G.L. FlashPCR: Revolutionising qPCR by Accelerating Amplification through Low ∆T Protocols. Int. J. Mol. Sci. 2024, 25, 2773.

Abstract

Versatility, sensitivity and accuracy have made the real-time polymerase chain reaction (qPCR) a crucial tool for research as well as diagnostic applications. However, for point-of-care (PoC) use traditional qPCR faces two main challenges: long run times mean results are not available for half an hour or more and the requisite high-temperature denaturation requires more robust and pow-er-demanding instrumentation. This study addresses both issues and revised primer and probe designs, modified buffers and low ∆T protocols which, together, speed up qPCR on conventional qPCR instruments and will allow the development of robust, point-of-care devices. Our approach, called "FlashPCR", also allows efficient reverse transcription as part of a one-step RT-qPCR pro-tocol, making it universally applicable for both rapid research and diagnostic applications.

Keywords

Reverse transcription; qPCR; COVID-19; molecular diagnostics; point-of-care

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

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