title: MRC Homo sapiens 6K
status: Public on Jan 17 2007
contact.city: Leicester
contact.first: Jerry
contact.middle: A
contact.last: Styles
contact.email: jas12@le.ac.uk
contact.state: Leicestershire
contact.address: Robert Kilpatrick Building, Univ. Leicester
contact.department: Cancer Studies and Molecular Medicine
contact.country: United Kingdom
contact.web_link: http://www.le.ac.uk/cm/rs/mrcmolend.html
contact.institute: University of Leicester
contact.phone: 441162231850
organism: Homo sapiens
manufacture_protocol: 5010 EST clones were obtained from Research Genetics (Human known gene set). The actual clones were obtained from the IMAGE collections held at the MRC Human Genome Mapping Project. These clones were sequence verified prior to arraying. All of the plasmid inserts were amplified by use of a universal primer set and a standard PCR mix (ABGene) employed. The products were all electrophoresed on an agarose gel and the gel images stored for future reference.
  All PCR products that failed an initial amplification or produced multiple bands were re-amplified using plasmid specific primers. The size of the insert sequence was determined from the agarose gel by comparison with ladder using MultiAnalyst software (BioRad). After PCR the reaction products were precipitated and prepared for array using the methods described by Eisen and DeRisi (DeRisi et al., 1997; Eisen et al., 1999).
  Arrays were printed on poly-L-lysine coated slides and UV crosslinked and blocked prior to use (DeRisi et al., 1997; Eisen et al., 1999). The arrays were printedusing an arrayer built essentially according to the Stanford designs and can be seen at the MRC toxicology website.  The centre to centre distance of the features was 210 um and each feature was 90-100 um in diameter