Total RNA was prepared from cross-sections of frozen uterus samples, using Nucleospin RNA II (Macherey-Nagel, Düren, Germany).
Label
biotin
Label protocol
2 micrograms of total RNA from each sample were used to prepare biotinylated fragmented cRNA according to the GeneChip® Expression Analysis Technical Manual (Rev. 5, Affymetrix Inc., Santa Clara, CA).
Hybridization protocol
Affymetrix GeneChip® expression arrays (Canine Genome 2.0 Array) were hybridized for 16 hours in a 45°C incubator, rotated at 60 rpm.
Scan protocol
According to the GeneChip® Expression Analysis Technical Manual (Rev. 5, Affymetrix Inc., Santa Clara, CA) the arrays were then washed and stained using the Fluidics Station 450 and finally scanned using the GeneChip® Scanner 3000 7G.
Description
Gene expression in uterine tissue from dog with pyometra disease.
Data processing
The data were analyzed with the robust multi-array average (RMA) method using default analysis settings.