Background: mixed genetic backgroud of C57Bl/6 and 129/SvJ Genotype: IRF6 null Age: Embryonic day 17.5
Skin from E17.5 mice was removed and flash frozen.
Total RNA was isolated as recommended (RNeasy, Qiagen, Valencia, CA), except that tissue was treated with proteinase K (20 mg/ml) for 20 minutes at 55°C.
Five micrograms of total RNA was processed for use on the microarray by using the Affymetrix GeneChip one-cycle target labeling kit (Affymetrix, Inc., Santa Clara, CA) according to the manufacturer’s recommended protocols
The resultant biotinylated cRNA was fragmented then hybridized to the GeneChip Mouse Genome 430 2.0 Array (45,000 probe sets used to analyze over 39,000 mouse transcripts and variants from over 34,000 well-characterized mouse genes; Affymetrix, Inc.). The arrays were washed and stained using the Affymetrix Model 450 Fluidics Station
GeneChips were scanned using the Affymetrix Model 3000 scanner
Gene expression data from E17.5 mouse skin, IRF6 null
Expression values were generated using the microarray suite (MAS) v5.0 software (Affymetrix, Inc)