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A comparison of the changes in rank order of individual miRNAs (Rank Difference AC-Con) and the log2 fold change from median normalized data [Median Normalized DCt (AC-Con)] is shown in Figure 1A, whereas a similar comparison of log2 fold change after Limma normalization to sno135 and miR-25 [DDCt AC-Con)] is shown in Figure 1B.
After log transformation of the data, we median normalized the data arrays in R language, using the Bioconductor biostatistical package (www.r-project.org/ and www.bioconductor.org/).
The microarray data were extracted using the Bioconductor limma package [39] and median normalized.
Log2 intensity ratios (M values) were then Median normalized to correct for differences in genomic DNA labelling efficiency between samples.
Data were analyzed with web-cellHTS2 Pelzz et al, 2010) to obtain plate-based median normalized z-scores for control and TRAIL-treated plates separately.
Signal intensities were collected with a GenePix 4000A Scanner and processed with GenePix software (Molecular Devices) and global median normalized with NOMAD (http://ucsf-nomad.sourceforge.net/).net/
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The signal intensities for each gene were then median-normalized and log2 transformed.
Heat maps were constructed using median-normalized data from each ProtoArray.
Log base 2 data were downloaded, transformed to linear, uploaded to GeneSpring GX 7.3 and then chips and genes were median-normalized.
Microarray data were median-normalized in Cy5 and Cy3 channels.
Every sample was manually median-normalized prior to analysis.
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