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Producing molecular biology reagents without purification.

Published version
Peer-reviewed

Type

Article

Change log

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

Keywords

COVID-19, COVID-19 Testing, Cameroon, Diagnostic Tests, Routine, Escherichia coli, Gene Expression, Geobacillus stearothermophilus, Ghana, Humans, Indicators and Reagents, Molecular Diagnostic Techniques, Plasmids, Real-Time Polymerase Chain Reaction, Recombinant Proteins, SARS-CoV-2, Synthetic Biology, Transformation, Bacterial, United Kingdom

Journal Title

PLoS One

Conference Name

Journal ISSN

1932-6203
1932-6203

Volume Title

16

Publisher

Public Library of Science (PLoS)
Sponsorship
NIBIB NIH HHS (R01 EB027202)
NIH HHS (1-R01-EB027202-01A1)