Question: What is the cutoff used for define high or low expression level of gene for survival analysis
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gravatar for mygamefun3
10 months ago by
mygamefun30
mygamefun30 wrote:

Hi everyone

In RNA-seq analysis, we need to separate samples into two groups for survival analysis. How can I define high level or low level for a gene according to counts or FPKM. Use median? average or quantile?

In TCGA or Oncomine, how are they define the cutoff for a gene ?

Thanks.

rna-seq • 1.1k views
ADD COMMENTlink written 10 months ago by mygamefun30

There's no definitive answer to your question. I would not advise going by the median or average. Quantile is a reasonable idea, or tertiles, with the higher third being regarded as "high expression".

An even better idea would be to convert your data to the Z scale, i.e., standard deviations from the mean, and then choose absolute 3, 4, 5, or 6 (3, 4, 5, or 6 standard deviations from the mean) as potential cut-offs. I trust that you have QC'd your data already and that low count transcripts have been removed.

ADD REPLYlink modified 9 months ago • written 10 months ago by Kevin Blighe31k

Thanks for your advice.

ADD REPLYlink written 10 months ago by mygamefun30

Hi Kevin,

I would like to look at survival of a specific gene between high and low expression tumor samples. For this, I want to divide tumor samples into high and low. I have counts data and transformed them into z-score.

Do you think the below one is right way to divide the samples?

event_rna <- t(apply(z_rna, 1, function(x) ifelse(x > 1.96,1,ifelse(x < -1.96,2,0))))

Or Should I follow this high and low samples separation

In that they used FPKM and took the median and based on that they separated samples into high and low.

Which one is the right solution?

ADD REPLYlink modified 3 months ago • written 3 months ago by Biologist90
1

Hey bro, There is no right or wrong. Your function looks okay!

ADD REPLYlink written 3 months ago by Kevin Blighe31k

Thank you very much Kevin.

ADD REPLYlink written 3 months ago by Biologist90
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