Evaluation of Normalization Methods for RNA-Seq Gene Expression Estimation.

Statistical inferences on RNA-Seq data, e.g., detecting differential gene expression, are meaningful only after proper normalization. However, there is no consensus for choosing a normalization procedure from among the many existing procedures. We evaluated several RNA-Seq normalization procedures by (1) correlating estimated RNA-Seq expression values to those of microarrays, (2) examining the concordance of stable and differential gene detection between the platforms, and (3) applying the procedures to simulated RNA-Seq data. Results suggested that RNA-Seq normalization procedures have little effect on both inter-platform gene expression correlation as well as inter-platform concordance of genes detected as stably or differentially expressed. However, the results of simulated analysis suggested that some normalization procedures are more robust to changes in distribution of differentially expressed genes. These results may provide guidance for selecting RNA-Seq normalization procedures.