title: UCHSC_Mouse_NIA15K
status: Public on Dec 02 2005
contact.city: Aurora
contact.first: Erik
contact.middle: James
contact.last: MacLaren
contact.email: erik.maclaren@uchsc.edu
contact.state: CO
contact.address: Mail Stop 8303, P.O. Box 6511
contact.department: Pharmacology
contact.country: USA
contact.institute: University of Colorado at Denver and Health Sciences Center
organism: Mus musculus
description: The National Institute of Aging Mouse 15K clone set was obtained from the
  Vanderbilt Microarray Shared Resource. Approximately 15,000 unique cDNA
  clones were derived from 52,374 ESTs from pre- and perimplantation embryos,
  E12.5 female gonad/mesonephros, and newborn ovary. Of the clones represented, 50%
  may be novel. There are 11,853 Unigene clusters represented and the set has been sequence verified. The average insert size is 1.5kb. For more information about the set
  go to http://lgsun.grc.nia.nih.gov/cDNA/15k.html .
manufacture_protocol: Plasmid DNAs were isolated from the 15K clone set using a high throughput Qiagen BioRobot 8000. Qiagen Plasmid prep reagents, and filter plates were used to extract and dispense the plasmid DNA into 96 well microtiter plates. The plasmids were assayed by gel electrophoresis for purity. A small random sampling of plasmids was done, and the inserts sequence verified by BLAST analysis. Plasmid inserts were amplified using universal primers, and an MJ Research Tetrad Thermocycler. The PCR products were purified using Millipore MultiScreen PCR plates. The PCR products were assayed by gel electrophoresis for purity and size. The PCR product set was very accurate, and clean. Occasionally, errors were found, and PCR products were re-amplified from the original clone set, or a replacement clone was ordered from Vanderbilt. The purified PCR products were dried down in a SC250DDA Speedvac Concentrator, and resuspended for distribution into 384 well microtiter plates for printing. Control PCR products including housekeeping gene, B-Actin, and several
  Arabidopsis genes were plated.  The PCR products and controls were resuspended in 3X SSC and printed with a GeneMachines OmniGrid 100 Arrayer onto Poly-L-Lysine coated microscope slides. The slides were blocked with succinic anhydride in 1-Methyl-2-Pyrrolidinone, and stored in a dessicator until hybridized.