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Producing molecular biology reagents without purification

Published version
Peer-reviewed

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Authors

Nguyen, Vylan 
Torres, Jose-Angel 
Kar, Shaunak 
Fadanka, Stéphane 

Abstract

We recently developed ‘cellular’ reagents–lyophilized bacteria overexpressing proteins of interest–that can replace commercial pure enzymes in typical diagnostic and molecular biology reactions. To make cellular reagent technology widely accessible and amenable to local production with minimal instrumentation, we now report a significantly simplified method for preparing cellular reagents that requires only a common bacterial incubator to grow and subsequently dry enzyme-expressing bacteria at 37°C with the aid of inexpensive chemical desiccants. We demonstrate application of such dried cellular reagents in common molecular and synthetic biology processes, such as PCR, qPCR, reverse transcription, isothermal amplification, and Golden Gate DNA assembly, in building easy-to-use testing kits, and in rapid reagent production for meeting extraordinary diagnostic demands such as those being faced in the ongoing SARS-CoV-2 pandemic. Furthermore, we demonstrate feasibility of local production by successfully implementing this minimized procedure and preparing cellular reagents in several countries, including the United Kingdom, Cameroon, and Ghana. Our results demonstrate possibilities for readily scalable local and distributed reagent production, and further instantiate the opportunities available via synthetic biology in general.

Description

Funder: Shuttleworth Foundation


Funder: Mozilla Foundation; funder-id: http://dx.doi.org/10.13039/501100012642

Keywords

Research Article, Biology and life sciences, Research and analysis methods, Medicine and health sciences, Engineering and technology

Journal Title

PLOS ONE

Conference Name

Journal ISSN

1932-6203

Volume Title

16

Publisher

Public Library of Science
Sponsorship
Welch Foundation (F-1654)
Bill and Melinda Gates Foundation (OPP1182120)
The National Institutes of Health (US) (1-R01-EB027202-01A1)