title: Indiana University/CGB-Daphnia-10K-Gen 3
status: Public on Mar 16 2011
contact.city: Notre Dame
contact.first: Jacqueline
contact.last: Lopez
contact.email: jacqueline.ann.lopez@gmail.com
contact.state: Indiana
contact.address: 019 Galvin Life Science Center
contact.department: Biology
contact.country: USA
contact.institute: University of Notre Dame
contact.phone: 574-631-1902
organism: Daphnia pulex
manufacture_protocol: 1. Lyophilized oligos are arrayed in Genetix 384-well polypropylene microarray plate. Each well contains XXX pmol of 70-mer oligo.
  2. Resuspend lyophilized oligos in 10ul of 150mM NaPO4, giving 30uM final concentration. 150mM NaPO4 is made from 150mM Na2HPO4 and 150mM NaH2PO4 in nuclease-free water and filtered with 0.45um Stericup Filter.
  3. Seal plates with Co-Star Corning Thermowell Aluminum Sealing tape (cat # 07200684)
  4. Centrifuge plate in Allegra 25R centrifuge at 2000 rcf for 1 minute.
  5. Incubate at 4 degrees Celcius for 16 hours to aid resuspension of DNA.
  6. Shake plates in flat bottom shaker at room temperature (15 -30 degrees Celcius) at 200 rpm for 60 mins.
  7. Centrifuge plate in Allegra 25R centrifuge at 2000 rcf for 1 minute.
  8. Store plates at 4 degrees Celcius until ready to print.
  9. Print 6 plates at a time to minimize evaporation; remove plates from 4 degree storage.
  10. Remove seal from plate. Place in appropriate plate cassette.
  11. Initiate print run method.
  12. After these 6 plates are printed, remove and reseal the plates. Store at 4 degrees Celcius. Repeat steps 9 and 10 until all desired plates are printed.
  13. After print run is complete, plates are stored at 4 degrees.