How to choose the samples you want to analyze?
New button in the Sample Selection panel. The selection of samples defines which data are used for the analysis.
Select particular conditions button to select all samples with a certain annotation, e.g. all data from a certain tissue type.
Note that you can select multiple tissues. When you select samples for use in the RefGenes tool, you have to focus on microarrays from samples that were collected in conditions similar to those in your qPCR experiment. Don’t make a too general selection, e.g. all human samples: you might end up with genes that are stable in most conditions but not in yours. Don’t make a very specific selection either, e.g. human heart samples from patients taking the same medication as yours. If you want to broaden your study later on with samples from other patients, your reference genes might not be valid anymore. It is recommended to select reference genes in the same organism and the same / a similar tissue type as the one that you used in your experiments.
This step is not essential, but it helps you to see whether your target gene(s) is (are) strongly or weakly expressed in the conditions of interest selected in STEP1. This allows you to search for candidate reference genes in a similar range of expression.
How to choose the genes you want to analyze?
New button in the Gene Selection panel.
The reference genes that are suggested by GeneVestigator have the following characteristics:
How to find the candidate reference genes?
Run button in the RefGenes tool. RefGenes will show the top 20 most stable genes with similar expression levels: