title: Rosenzweig_E.coli_4290_v1.0
status: Public on Jul 22 2009
contact.city: Missoula
contact.first: Frank
contact.last: Rosenzweig
contact.email: Frank.Rosenzweig@mso.umt.edu
contact.state: MT
contact.address: 32 Campus Drive, HS 104
contact.department: Division of Biological Sciences
contact.country: USA
contact.institute: University of Montana
organism: Escherichia coli str. K-12 substr. MG1655
description: Corning Gaps II aminosilane coated slides
manufacture_protocol: Microarrays were produced using full-length open reading frame PCR products generated with the Sigma-Genosys ORFmers primer set and reaction conditions and cycling parameters recommended by the manufacturer (Sigma-Genosys, The Woodlands, TX). This set contains primer pairs for all 4290 known and hypothetical ORFs in E. coli K12 MG1655. PCR reactions were repeated and pooled as necessary to obtain at least 3 micrograms of DNA and pooled reactions were ethanol precipitated, resuspended and further purified using a Qiagen MinElute96 UF PCR purification kit. Purified PCR products were run on agarose gels for quantification and to verify PCR product size. Products were standardized to each contain 2 µg, dried, and dissolved in 10 µl 3X SSC for printing. Arrays were printed onto Corning Gaps II aminosilane-coated slides using a 48-pin Stanford-UCSF style arrayer at the Stanford Functional Genomics Facility (Stanford, CA).