Normalization to Reference Genes
To normalize the expression of target genes to the expression of reference genes, select Normalization with reference gene(-s) in the Pre-processing menu in the Data editor to open a dialog box.
Select the gene to normalize in the left panel, select the appropriate reference gene(s) in the right panel, and press Select. Repeat this for all target genes that should be normalized so that all genes that should be normalized have type Norm. (normalized) in the left panel, and all reference genes type Ref. (reference). It is possible to select multiple genes in either panel (hold down Ctrl or Shift while selecting), and you can select different reference genes for different target genes. When you have defined all target and reference genes relationships, press Apply to execute normalization using the following equation.
GenEx will color the normalized columns (genes) yellow and remove the reference genes from the sheet. Any correction for PCR efficiency should be done before normalizing with reference genes. If your target genes are expressed at very different levels, consider using different reference genes for each target gene, so that the reference gene has matching expression levels. A tip is to use the Undo function to create and upload different data files to your project in the Control panel. Normalize the target genes with different reference genes or different number of reference genes to test the effect of normalization on the results of the analyses.