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[As a service to our readers, we will be gathering and reprinting notices posted on laboratory doors at the Massachusetts Institute of Technology. The following notice was found affixed to a door in the main biology building on the campus of MIT. It appears below unaltered. (Note: if you have notices that you have found posted to doors at MIT, please email them to mcsweeneysmail@yahoo.com with "MIT Doors" in the Subject header.)]

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E. coli ORFmers

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. Genosys Biotechnologies, Inc. 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.genetics.wisc.edu/). The PCR primer pairs were designed with the following features:

All upstream or "amino-terminal" primers contain an "ATG" start codon (A-primer).

All downstream or "carboxyl-terminal" primers contain a "TAA" stop codon (C-primer).

A Sap I restriction site has been incorporated into a 5' "adaptamer" sequence in all primers. The "adaptamer" sequence is 13 bases in length. 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.1

E. coli ORFmer Pricing:

$20 per PCR primer pair for orders of 1-20 pairs (2,000 pmol of A- and C-primers in individual tubes)

$12 per PCR primer pair for orders of 21 or more pairs (2,000 pmol of A- and C-primers in individual tubes).

$850 per 96-well plate (2,000 pmol of A- and C-primers combined per well {4000 pmol total}, 96 primer pairs).

Please contact your account representative if you are interested in purchasing a complete set of primers in tube or micro-plate formats (45 micro-plates, 4,290 PCR primer pairs).

1. Blattner, F.R., Plunkett, G., Bloch, C.A., Perna, N.T., Burland, V., Riley, M., Collado-Vides, J., Glasner, J.D., Rode, C.K., Mayhew, G.F., Gregor, J., Davis, N.W., Kirkpatrick, H.A., Goeden, M.A., Rose, D.J., Mau, B. and Shao, Y. The complete genome sequence of Escherichia coli K-12. Science 277:1453-1462 (1997).

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