Total RNA was isolated using Qiagen miReasy Mini kit TRI Reagent (Qiagen, Hilden, Germany) according to the manufacturer’s protocol for small-RNA. Then small RNA was cleaned up using the Rneasy MinElute Clean up kit (Qiagen, Hilden, Germany).
Label
biotin
Label protocol
Biotinylated cRNA were prepared from miRNA according to FlashTag Biotin RNA Labeling Kit for Affymetrix GeneChip miRNA arrays (Genisphere, Hatfield, PA, USA) according to the manufacturer’s recommendations Affymetrix GeneChip miRNA Array protocols (Affymetrix, Santa Clara, CA, USA)
Hybridization protocol
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 42C on Affymetrix GeneChip miRNA arrays. GeneChips were washed and processed to detect biotin-containing transcripts by Streptavidin-PE (Phycoerythrin) conjugate.
Scan protocol
Scanning using the GeneChip scanner 3000 were performed according to Affymetrix protocols
Description
Musculus longissimus dorsi tissue from adult pig taken 24h before slaughter miRNA expression data from skeletal muscle of adult pig breed (Duroc).
Data processing
Data were analysed with Expression Console and quantify by med polish
