title: Duke MicroRNA microarray
status: Public on Apr 04 2009
contact.city: Durham
contact.first: Jen-Tsan
contact.last: Chi
contact.email: jentsan.chi@duke.edu
contact.state: NC
contact.address: 101 Science Drive, DUMC 3382
contact.department: Molecular Genetics and Microbiology
contact.country: USA
contact.institute: Duke University
contact.phone: 919-668-4759
organism: Caenorhabditis elegans
  Drosophila melanogaster
  Danio rerio
  Homo sapiens
  Mus musculus
  Rattus norvegicus
manufacture_protocol: 1.Oligos are arrayed in Greiner 384-well flat-bottom plates. Each well contains 600 pmol of 70-mer oligo.
  2. Resuspend oligos in water to 20 uM and rearray 5 µL into 384-well, Genetix polystyrene V-bottom plates (cat# X6004).
  3. Allow Genetix plates to dry through passive water evaporation in a protected environment (e.g., chemical hood).
  4. Before printing, add 5 µL of 1X Printing Buffer to each well. This can be done the night before a print run is started.
  5. Seal plates with Corning seals.
  6. Incubate at 37°C for 30 minutes to aid resuspension of DNA.
  7. Shake plates near maximum rotational speed on flat-bed shaker for 1 minute.
  8. Centrifuge plates at 2000 rpm for 3 minutes.
  9. Remove seals and cover with plate lids. Place in appropriate location of plate cassette. This should be done with first plates just before print run is started to minimize evaporation time before printing. For second and third cassettes, wait until 30
  10. Make sure plates rest behind both holding clips in the cassettes. Push plates back into the cassettes as far as they will go, putting them in the proper position for the server arm.
  11. After the print run is completed, allow plates to dry through passive evaporation in a protected environment.
  12. For each subsequent preparation of these plates for a print run, add water to the wells instead of sodium phosphate buffer. The amount of water should be decreased by 0.25 µL per print run, as this is the amount drawn up by the pin capillary during ea