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From Cells to Cell-Free Protein Synthesis within 24 Hours Using Cell-Free Autoinduction Workflow
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Journal JoVE Bio-ingénierie
From Cells to Cell-Free Protein Synthesis within 24 Hours Using Cell-Free Autoinduction Workflow

From Cells to Cell-Free Protein Synthesis within 24 Hours Using Cell-Free Autoinduction Workflow

DOI:
10.3791/62866-v

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06:08 min

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July 22, 2021

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, , ,
1Department of Chemistry and Biochemistry,California Polytechnic State University San Luis Obispo, 2Center for Application in Biotechnology,California Polytechnic State University San Luis Obispo

Chapitres

  • 00:05Introduction
  • 00:36Media Growth
  • 01:35Cell Harvest
  • 02:34Extract Preparation
  • 03:43Cell-Free Protein Synthesis and Quantification of the Reporter Protein, Super Folder Green Fluorescent Protein (sfGFP)
  • 04:46Results: Comparison Between Cell Extracts Obtained from Cell-Free Autoinduction Media Grown to Optical Densities of 2.5 and 10
  • 05:27Conclusion

Summary

Traduction automatique

Traduction automatique

This work describes the preparation of cell extract from Escherichia coli (E. coli) followed by cell-free protein synthesis (CFPS) reactions in under 24 hours. Explanation of the cell-free autoinduction (CFAI) protocol details improvements made to reduce researcher oversight and increase quantities of cell extract obtained.

Tags

Keywords: Cell-free Protein SynthesisCell-free AutoinductionIn Vitro Protein SynthesisCell-free ExpressionEnergy MetabolismE. Coli BL21 StarS30 BufferCell Extract Preparation

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