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Producing molecular biology reagents without purification
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 instr...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8168896/ https://www.ncbi.nlm.nih.gov/pubmed/34061896 http://dx.doi.org/10.1371/journal.pone.0252507 |
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author | Bhadra, Sanchita Nguyen, Vylan Torres, Jose-Angel Kar, Shaunak Fadanka, Stéphane Gandini, Chiara Akligoh, Harry Paik, Inyup Maranhao, Andre C. Molloy, Jenny Ellington, Andrew D. |
author_facet | Bhadra, Sanchita Nguyen, Vylan Torres, Jose-Angel Kar, Shaunak Fadanka, Stéphane Gandini, Chiara Akligoh, Harry Paik, Inyup Maranhao, Andre C. Molloy, Jenny Ellington, Andrew D. |
author_sort | Bhadra, Sanchita |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8168896 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-81688962021-06-11 Producing molecular biology reagents without purification Bhadra, Sanchita Nguyen, Vylan Torres, Jose-Angel Kar, Shaunak Fadanka, Stéphane Gandini, Chiara Akligoh, Harry Paik, Inyup Maranhao, Andre C. Molloy, Jenny Ellington, Andrew D. PLoS One Research Article 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. Public Library of Science 2021-06-01 /pmc/articles/PMC8168896/ /pubmed/34061896 http://dx.doi.org/10.1371/journal.pone.0252507 Text en © 2021 Bhadra et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Bhadra, Sanchita Nguyen, Vylan Torres, Jose-Angel Kar, Shaunak Fadanka, Stéphane Gandini, Chiara Akligoh, Harry Paik, Inyup Maranhao, Andre C. Molloy, Jenny Ellington, Andrew D. Producing molecular biology reagents without purification |
title | Producing molecular biology reagents without purification |
title_full | Producing molecular biology reagents without purification |
title_fullStr | Producing molecular biology reagents without purification |
title_full_unstemmed | Producing molecular biology reagents without purification |
title_short | Producing molecular biology reagents without purification |
title_sort | producing molecular biology reagents without purification |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8168896/ https://www.ncbi.nlm.nih.gov/pubmed/34061896 http://dx.doi.org/10.1371/journal.pone.0252507 |
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