Dear all,

I am analysing qPCR data from the Exiqon where I have one card per sample,
in each card I have one observation for each miRNA. I have in total 8 cards,
2 for treatment 1, 3 for treatment 2 and 3 for treatment 3. Each card has
one endogenous gene, which I wouldn't like to use to normalize Ct values
because is being affected by the type of treatment. So I would like to use
scale.rank.
I am getting the following error:

sr.norm <- normalizeCtData(raw.cat, norm = "scale.rank")
Error in smooth.spline(ref[i.set], data[i.set]) :
  need at least four unique 'x' values

Does this mean I don't have enough replicates?

thanks for the help

Andreia

-- 
--------------------------------------------
Andreia J. Amaral
Unidade de Imunologia Clínica
Instituto de Medicina Molecular
Universidade de Lisboa
email: andreiaamaral@fm.ul.pt
          andreia.fonseca@gmail.com

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