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E. coli S30 Extract System for Circular DNA

A Powerful Tool for Identifying and Characterizing Polypeptides

  • Reduced chance of expressed proteins degrading
  • Includes all components needed for coupled transcription/translation
  • Produces very low levels of endogenous proteins, thus low backgrounds

Size

Catalog number selected: L1020

$ 458.00 Your price: Log In

E. coli S30 Extract System for Circular DNA
30 reactions
$ 458.00
Your price: Log In

The E. coli S30 Extract for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or λ vectors, providing a powerful tool for identifying and characterizing polypeptides. The investigator needs only to supply the cloned DNA containing the appropriate prokaryotic promoter and ribosome binding sites. The S30 Extract for Circular DNA Templates is prepared by modifications of the method described by Zubay from an E. coli strain B deficient in OmpT endoproteinase and lon protease activity. This results in a greater stability of expressed proteins that would otherwise be degraded by proteases if expressed in vivo. The S30 in vitro system also allows higher expression levels of proteins that are normally expressed at low levels in vivo due to the action of host-encoded repressors.

For more information, see the Protocols & Applications Guide.

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

Applications

  • Regulation of prokaryotic gene expression.
  • Identification of cloned DNA gene products or mapping of polypeptides to defined DNA fragments.
  • Rapid screening for translation from eukaryotic or prokaryotic DNAs cloned in vectors designed for high expression in E. coli.
  • Screening of compounds for inhibition of bacterial translation.
References
  1. Zubay, G. (1973) Ann. Rev. Genet. 7, 267–87.
  2. Zubay, G. (1980) Meth. Enzymol. 65, 856–77.
  3. Pratt, J.M. (1984) In: Transcription and Translation, Hames, B.D. and Higgens S.J., eds., IRL Press, Oxford, 179.
  4. Studier, F.W. and Moffatt, B.A. (1986) J. Mol. Biol. 189, 113–30.
  5. Collins, J. (1979) Gene 6, 29–42.

Protocols

Specifications

What's in the box?

Item Part # SizeConcentrationAvailable Separately

Amino Acid Mixture Minus Cysteine

L447A 1 x 175μl1mM View Product

S30 Extract, Circular

L464A 3 x 150μl

Luciferase Assay Reagent

L486A 1 x 250μl

Luciferase Dilution Reagent

L487A 1 x 1ml

pBESTluc™ DNA, Circular (Control)

L492A 1 x 10μg

S30 Premix without Amino Acids

L512A 1 x 750μl

Amino Acid Mixture Minus Leucine

L995B 1 x 175μl1mM View Product

Amino Acid Mixture Minus Methionine

L996B 1 x 175μl1mM View Product

SDS

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Use Restrictions

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

Storage Conditions

AA

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