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At the family level, also a significant dose responsive increase in microarray spot intensity was detected (with lower sensitivity).
Microarray spot intensity and quality data were extracted from the scanned images using the BlueFuse software version II (BlueGnome).
Microarray spot intensity signals were acquired by using Axon GenePix Pro 3.0 software package (Axon Instruments Inc).
Following strict quality assessment procedures in both techniques, (RNA quality, cross-hybridisation of microarray probes, microarray spot intensity, qPCR primer design, and PCR efficiency) and data filtering after normalization (cut-offs for fold-changes and low microarray spot intensity) resulted in high correlation between qPCR and microarray data (Additional file 9) [ 60, 61].
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With cDNA microarray, spot intensities are provided for all genes printed on the microarray slides, regardless of whether the genes are expressed or not.
We checked that mRNA as a proportion of total RNA (estimated by summation of all microarray spot intensities) did not differ between optimal growth conditions and isoleucine starvation.
It was previously found that the average expression level of 258 oocyte-expressed genes was lower on the X chromosome than other chromosomes, based on microarray spot intensities [ 48].
For each individual 8-mer, we derive both E-scores (which represent the relative rank of microarray spot intensities, and range from −0.5 to +0.5 [ Berger et al., 2006]) and Z-scores (which scale approximately with binding affinity [ Badis et al., 2009]).
For the determination of the global A. gambiae female salivary gland transcriptome, the microarray spot hybridization fluorescence intensities were used as an indicator of transcript abundance/expression level, since each spot contained a similar amount of nucleic acid.
Image analysis was carried out using the software Spot, a software package for the analysis of microarray images to extract spot intensity data from 16-bit tiff image files.
GenePix Pro 6.0 software was used to analyze the spot intensity of microarray data.
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