title: Edgerton Lab Candida albicans SC5314 6k array_ORF6
status: Public on Mar 05 2007
contact.city: Buffalo
contact.first: Wansheng
contact.last: Li
contact.state: NY
contact.address: 3435 main st.
contact.department: Oral Biology
contact.country: USA
contact.institute: University at Buffalo
organism: Candida albicans SC5314
manufacture_protocol: C. albicans ORFs were obtained
  from assembly 6 of the C. albicans genome sequence
  (http://www-sequence.stanford.edu/group/candida). ORFs encoding proteins greater than 150 aa and that started with an ATG codon were
  included in the array. ORFs were sorted into two categories: those
  that were unique by BLAST comparisons, and those that were likely
  to represent a group of closely related genes or a gene family.
  Primers to amplify each of the unique ORFs were designed using
  PRIMER3 software (http://frodo.wi.mit.edu/primer3/primer3_code.
  html). Primers for the gene family ORFs were designed by hand and
  checked using the PRIMER3 program. Primers were designed to have
  a Tm of approximately 60 uC and to amplify a region of approximately
  300 bp. When possible, amplification products were targeted
  to the 39 end of the coding region.
  Two separate rounds of PCR were conducted with each primer pair
  and C. albicans SC5314 genomic DNA as the template. PCR products
  from the reactions were combined and checked for quality by
  agarose gel electrophoresis. PCR products were purified using the MultiScreen-PCR 96-well filtration system (Millipore), eluted in water,
  dried in a SpeedVac (ThermoLabs) and resuspended at approximately
  0.1ug/ul in 3X SSC (0.45 M NaCl, 0.045 M sodium
  citrate). A total of 6392 DNA samples were spotted onto SuperAmine
  Substrate slides (Telechem) at Microarrays Inc. Controls included
  Arabidopsis thaliana spot report genes and mammalian DNA
  fragments (Stratagene), C. albicans genes of known expression profiles,
  and buffer blanks.