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JoVE Journal

Bioengineering

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

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06:08 min

July 22nd, 2021

July 22nd, 2021

4,076 Views

0:05

Introduction

0:36

Media Growth

1:35

Cell Harvest

2:34

Extract Preparation

3:43

Cell-Free Protein Synthesis and Quantification of the Reporter Protein, Super Folder Green Fluorescent Protein (sfGFP)

4:46

Results: Comparison Between Cell Extracts Obtained from Cell-Free Autoinduction Media Grown to Optical Densities of 2.5 and 10

5:27

Conclusion

Transcription

The cell-free autoinduction method reduces researcher oversight and time needed to produce functional cell extracts. This simplifies previous in vitro protein synthesis protocols that were more time-consuming and intensive. This technique is less

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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.

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