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E. coli T7 S30 Extract System for Circular DNA

L1130_E- coli T7 S30 Extract System for Circular DNA_3
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Create Radiolabeled Protein for Transcription/Translation Studies

  • Translate using clones with a T7 promoter and a ribosome binding site; no E. coli promoter required
  • Reduce the chance of expressed proteins degrading
  • Includes all components needed for coupled transcription/translation

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Catalog number selected: L1130

$ 595.00
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E. coli T7 S30 Extract System for Circular DNA
30 reactions
$ 595.00
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Translation from Any Clone with a T7 Promoter

The E. coli T7 S30 Extract System for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or λ vectors containing a T7 promoter by providing an extract that contains T7 RNA polymerase for transcription and all components needed for translation. The investigator only supplies cloned DNA containing a T7 promoter and a ribosome binding site. This product 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 greater stability of expressed proteins that would otherwise be degraded by proteases if expressed in vivo.

The E. coli T7 S30 Extract System for Circular DNA enables you to translate using any clone that has a T7 promoter and a ribosome binding site; No E. coli promoter required. Proteins expressed with this system demonstrate reduced tendency to degrade and have shown very low levels of endogenous protein.

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
  • Synthesis of radiolabeled protein.
  • Verification of cloned gene expression before in vivo protein expression in E. coli.
  • Use of protein in functional studies of transcription and translation.
  • Use of protein as a tracer in protein purification.
  • Incorporation of unnatural amino acids into proteins for structural studies.

References

  1. Nevin, D.E. and Pratt, J.M. (1991) FEBS Lett. 291, 259–63.
  2. Zubay, G. (1973) Ann. Rev. Genet. 7, 267–87.
  3. Zubay, G. (1980) Meth. Enzymol. 65, 856–77.
  4. Studier, F.W. and Moffatt, B.A. (1986) J. Mol. Biol. 189, 113–30.
  5. Pratt, J.M. (1984) In: Transcription and Translation, Hames, B.D. and Higgens S.J., eds., IRL Press, Oxford, 179.

Specifications

What's in the box?

Item Part # Size Concentration Available Separately

T7/S30 Extract for Circular DNA

L114A 3 × 150μl

Amino Acid Mixture Minus Cysteine

L447A 1 × 175μl 1mM

S30 Premix without Amino Acids

L512A 1 × 750μl

Amino Acid Mixture Minus Leucine

L995B 1 × 175μl 1mM

Amino Acid Mixture Minus Methionine

L996B 1 × 175μl 1mM View Product

PinPoint™ Xa Control Vector

V204A 1 × 5μg

Certificate of Analysis

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Use Restrictions

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

Storage Conditions

AA

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