title: An oligonucleotide microarray for mouse imprinted genes profiling
status: Public on Apr 01 2012
contact.city: Paris
contact.first: Jean-Philippe
contact.last: Jais
contact.email: jean-philippe.jais@parisdescartes.fr
contact.address: 149 rue de Sevres
contact.country: France
contact.institute: Université Paris Descartes - Hopital Necker-Enfants Malades
organism: Mus musculus
  Rattus norvegicus
manufacture_protocol: Specific 50-mer oligonucleotides synthesized by Operon were resuspended in 150 mM phosphate buffer pH 8.3 at a concentration of 20 micromoles. For each gene, two different oligonucleotide probes, one corresponding to the 3’ non-coding region and one corresponding to the coding region were designed. The resulting application targeted microarray contained 992 gene-specific oligonucleotide probes generated from 63 imprinted genes and 15 genes encoding proteins involved in epigenetic marking (BORIS, brother of regulatory imprinting sequence, 5 DNMT, DNA methyltransferases and 9 HDAC, histone deacetylases), as well as 256 genes encoding proteins involved in neurotransmission and 157 genes encoding proteins involved in energy homeostasis. These sequences correspond to mouse imprinted genes listed at http://www.geneimprint.com or reported in published papers. Probes were spotted on NHS-PEG (N-hydroxysuccinimide-polyethyleneglycol-vinylsulfone) activated glass slides (Genescore) (Pintchovski et al., 1979). Each probe was spotted in duplicate (at least) on each slide. The various oligonucleotide solutions were applied on the slides by contact printing, using a Biorobotics Microgrid II apparatus (Genomic Solutions, Ann Arbor, MI) with one BioRobotics MicroSpot 2500 pin. The oligonucleotide spots were 80–100 micrometers in diameter, and the center-to-center distance between adjacent spots was 220 micrometers. Each slide contained one array in a conformation of four rows and two columns. During spotting, relative humidity was maintained at 50%. After spotting, the arrays were left in the humidity chamber for a short period and were then stored in a desiccation chamber at 20 ° C until use.