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For analysis of gene expression patterns in the 64-sample set, the signal value for each probe in each sample was calculated as the log-transformed ratio of normalized intensity versus the background (common for all microarray data after normalization).
The microarray data after normalization were subjected to PCA analysis to visualize gene expression differences between control and treated animals.
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The assumption of normality is rarely satisfied in microarray data even after normalization.
After microarray data normalization, Principal Component Analysis was performed on the average gene expression profiles of the experimental groups.
Therefore, similar to the normalization of microarray data, quantile normalization on the multiplex RT-PCR results was performed.
Since the normalization procedure greatly changes the microarray data, different normalization procedures may lead to very different clustering results.
While microarray data normalization strategies are well established, an ideal method for RNA-seq normalization has yet to be defined.
After scanning, the microarray data were normalized using quantile normalization in the R language environment (version 2.8.1, available at http://www.r-project.org/).org/
Quackenbush, J. Microarray data normalization and transformation.
Commonly, microarray data normalization methods assume that relatively few transcripts change from sample to sample [36].
Our data integration approach includes microarray data normalization, transformation, and quality control.
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