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Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology
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Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology

DOI:
10.3791/50762-v

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16:11 min

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September 16, 2013

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, , , , ,
1Department of Biology,California Institute of Technology, 2Department of Bioengineering,California Institute of Technology, 3Synthetic Biology Center, Department of Bioengineering,Massachusetts Institute of Technology, 4School of Physics and Astronomy,University of Minnesota

Chapters

  • 00:05Title
  • 01:41Cell Lysis
  • 06:29Extract Clarification
  • 08:32Experimental Execution of a TX-TL Reaction
  • 10:40Results: Assessment of TX-TL Cell-free Expression System
  • 15:56Conclusion

Summary

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This five-day protocol outlines all steps, equipment, and supplemental software necessary for creating and running an efficient endogenous Escherichia coli based TX-TL cell-free expression system from scratch. With reagents, the protocol takes 8 hours or less to setup a reaction, collect, and process data.

Tags

Keywords: Cell-free Expression SystemEscherichia ColiTX-TLTranscription-translationSynthetic BiologyProtein ExpressionIn VitroEndogenousT7 RNA PolymeraseLac/tet RepressorsMRNA DegradationDNA DegradationBiomolecular Breadboard

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