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Escherichia coli-Based Cell-Free Protein Synthesis
 

Escherichia coli-Based Cell-Free Protein Synthesis: Protocols for a robust, flexible, and accessible platform technology

Article DOI: 10.3791/58882-v • 09:45 min • February 25th, 2019
February 25th, 2019 •
*1,2, *2,3, 4,5,6,7, 2,3, 2,3
1Department of Biological Sciences, California Polytechnic State University, San Luis Obispo, 2Center for Applications in Biotechnology, California Polytechnic State University, San Luis Obispo, 3Department of Chemistry and Biochemistry, California Polytechnic State University, 4Department of Chemical and Biological Engineering, Northwestern University, 5Chemistry of Life Processes Institute, Northwestern University, 6Center for Synthetic Biology, Northwestern University, 7Interdisciplinary Biological Sciences Program, Northwestern University
* These authors contributed equally

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This protocol details the steps, costs, and equipment necessary to generate E. coli-based cell extracts and implement in vitro protein synthesis reactions within 4 days or less. To leverage the flexible nature of this platform for broad applications, we discuss reaction conditions that can be adapted and optimized.

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Escherichia Coli Cell-free Protein Synthesis Protocol Self-reprotein Synthesis Accessible Platform Technology Speed Cost-effectiveness Reaction Set Up Functional Genomics High-throughput Testing Biosensors Educational Kits Metabolic Engineering Genetic Code Expansion Sonication Media Preparation Culture Preparation Centrifuge Bottle Ice Water Bath OD600 Measurement Centrifugation
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