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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.
  • Bhadra S; Department of Molecular Biosciences, College of Natural Sciences, The University of Texas at Austin, Austin, Texas, United States of America.
  • Nguyen V; Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, Texas, United States of America.
  • Torres JA; Freshman Research Initiative, DIY Diagnostics Stream, The University of Texas at Austin, Austin, Texas, United States of America.
  • Kar S; Freshman Research Initiative, DIY Diagnostics Stream, The University of Texas at Austin, Austin, Texas, United States of America.
  • Fadanka S; Department of Molecular Biosciences, College of Natural Sciences, The University of Texas at Austin, Austin, Texas, United States of America.
  • Gandini C; Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, Texas, United States of America.
  • Akligoh H; Mboalab Biotech, Yaoundé, Cameroon.
  • Paik I; Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge, United Kingdom.
  • Maranhao AC; Hive Biolab, Kumasi, Ghana.
  • Molloy J; Department of Molecular Biosciences, College of Natural Sciences, The University of Texas at Austin, Austin, Texas, United States of America.
  • Ellington AD; Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, Texas, United States of America.
PLoS One ; 16(6): e0252507, 2021.
Article in English | MEDLINE | ID: covidwho-1388918
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Diagnostic Tests, Routine / Real-Time Polymerase Chain Reaction / COVID-19 Testing / SARS-CoV-2 / COVID-19 / Indicators and Reagents Type of study: Diagnostic study / Observational study Limits: Humans Country/Region as subject: Africa / Europa Language: English Journal: PLoS One Journal subject: Science / Medicine Year: 2021 Document Type: Article Affiliation country: JOURNAL.PONE.0252507

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Diagnostic Tests, Routine / Real-Time Polymerase Chain Reaction / COVID-19 Testing / SARS-CoV-2 / COVID-19 / Indicators and Reagents Type of study: Diagnostic study / Observational study Limits: Humans Country/Region as subject: Africa / Europa Language: English Journal: PLoS One Journal subject: Science / Medicine Year: 2021 Document Type: Article Affiliation country: JOURNAL.PONE.0252507