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Cell-Free Protein Synthesis from Exonuclease-Deficient Cellular Extracts Utilizing Linear DNA Templates
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Cell-Free Protein Synthesis from Exonuclease-Deficient Cellular Extracts Utilizing Linear DNA Templates

Article DOI: 10.3791/64236-v • 08:11 min • August 9th, 2022
August 9th, 2022 •
*1, *1, 1, 2,3, 4, 1
1INRAe, AgroParisTech, Micalis Institute, Université Paris-Saclay, 2Helmholtz Institute for RNA-based Infection Research (HIRI), Helmholtz-Centre for Infection Research (HZI), 3Medical Faculty, University of Würzburg, 4Centre de Biochimie Structurale, INSERM U1054, CNRS UMR 5048, University of Montpellier
* These authors contributed equally

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Presented here is a protocol for the preparation and buffer calibration of cell extracts from exonuclease V knockout strains of Escherichia coli BL21 Rosetta2 (ΔrecBCD and ΔrecB). This is a fast, easy, and direct approach for expression in cell-free protein synthesis systems using linear DNA templates.

Tags

Cell-Free Protein Synthesis Exonuclease-Deficient Cellular Extracts Linear DNA Templates Cloning Chemical Modifications Protective Supplements Self-reporting Synthesis Extracts Prototyping BL21 Rosetta-2 Delta RecBCD Strain YTP Agar Plate Chloramphenicol Incubate RPM Shaking Subculture Optical Density
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