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The pooling is necessary because it ensures that sufficient salivary mRNA can be obtained for microarray analyses.
69% of the PUM1 mRNA targets (p<10−100) and 74% of the PUM2 targets (p<10−100) contained the consensus motif in the 3'-UTR, which represents a striking enrichment compared to its genome-wide occurrence in 3'-UTRs (20% of all ENSEMBL annotated genes; 22% of all genes for which data could be obtained for microarray cDNA probes).
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Liver biopsies were obtained for microarray and quantitative real-time PCR analyses of gene expression.
At each of the time-points at which RNA was obtained for microarray experiments, we quantified the expression of at least one transcript.
From the initial set of 36,230 blastula stage ESTs, a non-redundant set of ∼6,800 annotated sequences was obtained for microarray analysis.
Mice fat-fed for 2 days, 8 days, 3 weeks and 15 weeks, and with age-matched controls, were killed and epididymal WAT was obtained for microarray gene expression studies, employing Affymetrix mouse genome 4302 microarray.
Three tissue cores measuring 1.0 mm from each block were obtained for tissue microarray (TMA) (Beecher Instruments, Sun Prairie, WI).
Two technical replicates were obtained for each microarray experiment.
Intensity data and Cy5/Cy3 ratio (raw) data were obtained for individual microarray features at each scan time using the Axon GenePix Pro software.
Correlation coefficient values of R 2 = 0.97 and 0.95 were obtained for comparisons between microarray and qPCR for the time points of 30 and 120 min, respectively, which indicated microarray data were of good quality.
Above-background quantitative signal values were obtained for the Agilent microarray.
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