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P9733

Sigma-Aldrich

ORFmer™ PCR Primer Pairs

Escherichia coli

form

lyophilized powder

storage temp.

2-8°C

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General description

E. coli ORFmers are PCR primer pairs that have been designed to amplify each open reading frame (ORF) in E. coli, from the start codon to the stop codon. Sigma-Genosys offers 4,290 primer pairs, representing all protein-coding genes. The E. coli ORFmers are the direct result of a collaboration with the E. coli Genome Group, headed by Dr. Fred Blattner at the Department of Genetics, University of Wisconsin-Madison (http://www.genome.wisc.edu).

Application

Specially designed primers for the optimal amplification of all recognized protein-coding genes from ten microorganisms. Amplification of the genes can be used to make gene arrays or can be cloned for further characterization.

Features and Benefits

  • All upstream or "amino-terminal" primers contain an "ATG" start codon (A-primer) and all downstream or "carboxyl-terminal" primers contain a "TAA" stop codon (C-primer).
  • All primers have a 13 base adaptamer sequence attached to the 5′ end.
  • A Sap I restriction site has been incorporated into a 5′ "adaptamer" sequence in all primers. The Sap I enzyme cleaves outside its recognition sequence, generating a PCR product with an 5′-ATG-3′ overhang at one end and a 5′-TTA-3′ overhang at the other end. This feature has been included to facilitate cloning into a vector with the corresponding sites.
  • A pair of "Universal Adaptamer" PCR primers are available to allow re-amplification of any ORF PCR product.
  • ORFmer Primers are sorted according to gene "Functional Group". Primers from any functional group may be ordered as individual tubes or in a 96-well micro-plate format. There are a total of 23 functional groups.
  • PCR primer pairs to amplify most or all open reading frames (ORFs) from the microbial genome.
  • Designed to amplify the portions of each ORF that show minimal homology to other ORFs in that genome.
  • Available as lyophilized primers, one pair per well in a 96-well plate format.
  • Primers contain a common 5′ tag sequence (adaptamer) to allow easy re-amplification with a single pair of universal adaptamer primers.
  • Restriction sites are incorporated into the adaptamer sequences for cloning and further characterization.

Packaging

The ORFmers are available as lyophilized products in microcentrifuge tubes or as opposing pairs in a 96-well plate format.

Certificate of Analysis

Certificate of Origin

A complete set of PCR primers for the E. coli genome.
Richmond, C., Blattner, F.R.
Genosys Origins, 1, 3-4 (1998)
F R Blattner et al.
Science (New York, N.Y.), 277(5331), 1453-1462 (1997-09-05)
The 4,639,221-base pair sequence of Escherichia coli K-12 is presented. Of 4288 protein-coding genes annotated, 38 percent have no attributed function. Comparison with five other sequenced microbes reveals ubiquitous as well as narrowly distributed gene families; many families of similar...

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