title: VMSR Mus 20K version 2
status: Public on Jul 12 2007
contact.city: Los Angeles
contact.first: Mark
contact.middle: H.
contact.last: Chin
contact.email: mchin@mednet.ucla.edu
contact.state: CA
contact.address: 650 Charles E. Young Dr
contact.department: Human Genetics
contact.country: USA
contact.institute: UCLA
contact.phone: (310)794-5711
organism: Mus musculus
manufacture_protocol: 1. Microarrays containing 22,172 cloned genes from the National Institute on Aging 15k cDNA clone set, mouse sequence-verified clones from Research Genetics and several control genes were generated by the Vanderbilt Microarray Shared Resource.
  2. Briefly, all cDNA clones were amplified (Hegde, P, et. al., Biotechniques, 29, 548-562.) and verified by gel electrophoresis.
  3. The dried PCR products were then resuspended in 40 ml VMSR buffer A [Vanderbilt Microarray Shared Resource, Nashville, TN] resulting in an average concentration of 400 – 700 mg/ml per product.
  4. The PCR products were then transposed to 384 well plates and robotically arrayed using a BioRobotics MicroGrid II microarray printing robot (Apogent Discoveries, Hudson, NH) onto amine-coated  glass slides (TeleChem International, Sunnyvale, CA).
  5. The microarrays were then crosslinked using  80mjoules of ultraviolet energy (Stratgene Stratalinker, LaJolla, CA).
  6. Following crosslinking, the arrays were baked for two hours at 70°C in a standard oven and stored under low humidity conditions until use.