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E. coli S30 Extract System for Linear Templates

E. coli S30 Extract System for Linear Templates
30 reactions
$ 493.00
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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, λPL (con) and λ-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.

Specifications

What's in the box?

Item Part # SizeConcentrationAvailable Separately

Amino Acid Mixture Minus Cysteine

L447A 1 × 175μl1mM View Product

S30 Extract, Linear

L480A 3 × 150μl

pBESTluc™ DNA, Linear (Control)

L485A 1 × 10μg

Luciferase Assay Reagent

L486A 1 × 250μl

Luciferase Dilution Reagent

L487A 1 × 1ml

S30 Premix without Amino Acids

L512A 1 × 750μl

Amino Acid Mixture Minus Leucine

L995B 1 × 175μl1mM View Product

Amino Acid Mixture Minus Methionine

L996B 1 × 175μl1mM View Product

SDS

Choose language:

Download SDSPDF (213 KB) – English (United States)

Use Restrictions

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

Storage Conditions

AA

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