title: DNA Chip Consortium Mouse Microarray v2.0
status: Public on Dec 20 2005
contact.city: Sakyo-ku, Kyoto-shi
contact.first: Akihiro
contact.last: Matsunaga
contact.email: a-matsunaga@mfour.med.kyoto-u.ac.jp
contact.state: Kyoto
contact.address: Yoshida-konoe-cho
contact.department: Medicine
contact.country: Japan
contact.institute: Kyoto University
organism: Mus musculus
description: The oligonucleotide microarray is composed of 21987 of 65 mer oligonucleotides from the Sigma Geneosys oligo DNA library.  Oligonucleotides were printed by Hitachi Software Engineering.
manufacture_protocol: Total RNA is extracted using TRI Reagent (Sigma) and amplified with AminoAllyl MessageAmp aRNA Amplification Kit (Ambion) according to the manufacturer’s protocol.  The amino allyl aRNA is labeled by coupling reaction with NHS ester fluorescent dye, Cy3 or Cy5.  Each labeled aRNA sample is diluted with 5 × Fragmentation Buffer (DNA Chip Consortium) and fragmented by boiling at 94 ºC for 15 min after.  Hybridization mixture is prepared using labeled and fragmented aRNA at 0.4 μg/μl in 5 × SSC, 0.5% SDS, 4 × Denhardt’s solution, and 20% hibridization solution (DNA Chip Consortium).  The hybridization mixture is denatured at 95 ºC for 2 min and then mixed with salmon sperm DNA at 100 µg/ml (Sigma) and 10% Formamide.  The mixture is hybridized for 16 hr at 60 ºC in a hybridization oven.  After hybridization, the microarray is washed with solution I (2 × SSC, 0.1% SDS), solution II (2 × SSC), and solution III (1 × SSC) for each 5 min at 30ºC.  After washing, the microarray is scanned.