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A more limited series of confirmatory relative gene expression experiments, focused specifically on those genes identified as being significantly up- or downregulated using the microarray procedure, were also performed.
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A calibration curve, consisting of several dilutions of cDNA of the reference used for the microarray procedure, was included for each gene to allow calculation of the relative starting quantities.
Microarrays are becoming less popular due to the decreasing costs of the RNA-seq methods [ 100], although one has to remember that some of the steps used in the microarray procedure, with their drawbacks and limitations, are also utilized in other techniques including the RNA-seq approaches [ 101– 101].
The microarray procedure used in this study was essentially as described in [ 14].
The microarray procedure was carried out as described previously.
Using the same microarray data screening procedure described for the glomerular compartment, only requiring five fold upregulation instead of four, fourteen genes were identified with elevated expression in medullary endothelial cells, as described below.
Microarray analysis was carried out using the recommended procedure supplied by Research Genetics Inc.
The correlation analysis between qPCR and microarray expression data was performed using the CORR procedure of SAS SAS Inst . Inc).
Hybridization cocktails were applied to Affymetrix Rat Gene 2.0 microarrays and processed in accordance with the manufacturer's recommended procedure using the GCS3000 microarray system (Affymetrix).
Hybridisation cocktails were applied to Affymetrix Rat Gene 2.0 microarrays, and processed in accordance with the manufacturer's recommended procedure using the GCS3000 microarray system (Affymetrix).
We generated microarray probes for squids, I. paradoxus, using the following procedure.
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