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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
Kapitel
- 00:04Titel
- 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
Denne protokol beskriver de skridt, omkostninger og nødvendige udstyr til at skabe E. coli-baseret celle ekstrakter og gennemføre in vitro- protein syntese reaktioner inden for 4 dage eller derunder. For at udnytte de fleksible karakter af denne platform for brede programmer, diskuterer vi reaktionsbetingelser, der kan tilpasses og optimeret.
