Thoracic aorta dissected, cut into 2mm rings, washed 12 times in serum free EBM (Cambrex BioScience) and cultured for 14 days in serum free-EBM, changing medium every third day.
Extracted molecule
total RNA
Extraction protocol
Four to Six individual rings combined, snap frozen in liquid nitrogen, crushed and extracted with Trizol reagent (Invitrogen) followed by RNAEasy Micro kit (Qiagen) purification with DNAse I treatment. Total RNA eluted into DEPC water.
Label
Phycoerythrin
Label protocol
Standard Affymetrix "two-cycle" target labeling
Hybridization protocol
Standard Affymetrix protocol
Scan protocol
Standard Affymetrix protocol
Description
RNA labeling, array hybridization, scanning, and initial data transformation done at the Center for Expression Arrays at the Univesity of Washington.
