title: Su Lab Mouse 23K cDNA microarray
status: Public on Feb 09 2009
contact.city: Bethesda
contact.first: Tomoshige
contact.last: Kino
contact.email: kinot@mail.nih.gov
contact.state: MD
contact.address: 10 Center Drive MSC 1109
contact.department: Program in Reproductive and Adult Endocrinology
contact.country: USA
contact.institute: National Institute of Child Health & Human Development
contact.phone: (301)496-6417
organism: Mus musculus
description: This array includes the NIA mouse 15K and 7.4K cDNA clones as well as 0.2K controls
manufacture_protocol: 1. cDNA clones of the mouse 23K genes are amplified by PCR using published methods (Su YA and Trent JM, in Methods in Molecular Medicine, B. J. Nickoloff, Ed. [The Human Press Inc., Totowa, NJ, 2001], pp. 15-29.)
  2. The PCR-amplified DNA fragments are arrayed in 96-well V-bottom plates (Matrix Cat#4914). Each well contains approximately 0.2 micrograms cDNA in 10 microlitters 3 X SSC
  3. Dry the DNA-containing solution via evaporation at room temperature in stainless drawers within a class 10,000 clean room the day before printing.
  4. Two hours before printing, add 10 µL of ultrapure water to each well.
  5. Seal plates with Corning seals.
  6. Incubate at room temperature for at least two hours to aid resuspension of DNA.
  7. Shake plates near maximum rotational speed on flat-bed shaker for 1 minute.
  8. Spin plates at 2000 rpm (Sorvall Super T21) at room temperature for 5 minutes.
  9. Remove seals and cover with plate lids. Place plates in the Server Arm cassettes of GeneMachine OmniGrid 100 at 70% humidity.
  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 (same as the step 3 above)
  12. For each subsequent preparation of these plates for a print run, the amount of ultrapure water added should be decreased by 0.25 µL per print run, as this is the amount drawn up by the pin capillary during each print cycle.