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Escherichia coli-Based Cell-Free Protein Synthesis: Protocols for a robust, flexible, and accessible platform technology
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
Chapters
- 00:04Title
- 00:50Cell Growth
- 03:30Crude Cell Extract Preparation
- 05:30Cell-Free Protein Synthesis Batch Format Reactions
- 06:25Quantification of the Reporter Protein, [sfGFP]
- 07:24Results: Modification and Optimization of the Cell-Free Platform Conditions
- 09:01Conclusion
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.
