Labeled cRNAs were fragmented to an average size of approximately 50 to 100 nucleotides in the presence of 10 mM zinc chloride and adding a hybridization buffer containing 1 M sodium chloride, 0.5 percent sodium sarcosine, 50 mM morpholino-ethane sulfonic acid (pH 6.5), and formamide (final concentration, 30 percent at 40°C); After hybridization, the slides were washed
Scan protocol
slides were scanned using a confocal laser scanner (Agilent Technologies, Palo Alto, CA). Fluorescence intensities of the scanned images were quantified, normalized, and balanced
Description
Samples from liver, hypothalamus, pancreatic islets, adipose (gonadal fat pads) and muscle (soleus and gastrocnemius) were acquired from C56Bl/6 and Btbr male mice, bred from in-house colonies at the University of Wisconsin Biochemistry Department. Individuals from each strain were compared to a common pool created from equal portions of RNA from 20 conspecific samples.
Data processing
Rosetta Resolver Ratio Experiment using Rosetta Resolver Agilent Error Model
