Question: Independent groups normalisation
1
gravatar for elb
21 months ago by
elb180
Torino
elb180 wrote:

Hi guys, I have a question about the normalization of RNA Seq data. I have 3 types of cells of which I would like to identify the differentially expressed genes. As reported in many papers and tutorials, data have to be normalized together. However, I'm afraid that the normalisation could affect the expression of the genes of one of the three groups for some reasons that I do not know perfectly. Is there a way to make the groups comparable after making a normalisation internal to each cell type?

Thank you in advance

rna-seq normalization • 394 views
ADD COMMENTlink modified 21 months ago by WouterDeCoster44k • written 21 months ago by elb180

Is there a way to make the groups comparable after making a normalisation internal to each cell type?

RPKM / FPKM data is derived from normalisation on a per sample basis; however, this then renders the samples incomparable via statistical analyses (but do not tell this to the 1000s of researchers who, despite this, go ahead and perform differential expression analysis with RPKM / FPKM data).

ADD REPLYlink modified 21 months ago • written 21 months ago by Kevin Blighe61k

Before trying to solve a problem, check if there is a problem. If you don't know beforehand if the normalization will work or not, why don't you first try and evaluate the results? Examine the diagnostic plots, the number and pattern of differentially expressed genes between each contrast, and so on.

ADD REPLYlink written 21 months ago by h.mon30k
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