E. coli S30 Extract System for Linear Templates

by Promega
Available SKUs: L1030 |
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Description

  • Overview
  • Protocols
  • Specifications
  • Resources
  • Successful Transcription/Translation with a Variety of Templates

    The E. coli S30 Extract System for Linear Templates is prepared using minor modifications of the protocol described by Lesley and colleagues and allows successful transcription/translation of linear DNA templates. The investigator need only provide linear DNA containing a prokaryotic E. coli-like promoter (such as lacUV5, tac, ?PL (con) and ?-PR). A ribosome binding site is required to direct the synthesis of proteins in vitro. In vitro-generated RNA from DNA templates lacking an E. coli promoter may also be used in this system, but protein yields will be decreased to 1–10% of that produced from linear DNA templates.

    The E. coli S30 Extract System for Linear Templates allows the use of many different templates, including DNA fragments, PCR-synthesized DNA, ligated overlapping oligonucleotides, in vitro-generated RNA and prokaryotic RNA. The use of this system reduces the chance of expressed proteins degrading.

    For more information, see the Protocols & Applications Guide.

    Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/custom/

    Applications

    • Identification of cloned or synthesized genes from linear or closed circular DNA.
    • Rapid structure mapping of genes without additional cloning steps.
    • Rapid verification of in vitro-generated mutations.
    • Synthesis of truncated gene products for functional domain mapping and epitope mapping studies.
    • Expression from in vitro-generated RNA for genes lacking an appropriate E. coli promoter.

    Reference

    1. Lesley, S.A. et al. (1991) J. Biol. Chem. 266, 2632–8.
  • Protocols

    Complete Protocol

    Download PDF

    E. coli S30 Extract System for Linear Templates Technical Bulletin

    PDF (767 KB)

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    LESS THAN -65C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

  • Resources

    Citations

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