E. coli S30 Extract System for Linear Templates
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/
- 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.
- Lesley, S.A. et al. (1991) J. Biol. Chem. 266, 2632–8.
E. coli S30 Extract System for Linear Templates Technical Bulletin
PDF (767 KB)
Certificate of AnalysisLookup Certificate of Analysis
LESS THAN -65C
For Research Use Only. Not for Use in Diagnostic Procedures.