Different Normalizations
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@julia-engelmann-559
Last seen 10.2 years ago
Hi, I am comparing different normalizations of Affymetrix microarrays: 1. expresso with rma, vsn, pmonly and medianpolish 2. calling rma and vsn with: eset=rma(Data); Set=vsn(eset); I know that medianpolish does log2 transformation and vsn does log transformation (base e), but when I compare the results, the difference between the two datasets is huge- not a matter of log-base. Shouldn't the results be very similar? Any help would be much appreciated. Thanks, Julia
vsn vsn • 1.7k views
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Susan Holmes ▴ 120
@susan-holmes-158
Last seen 10.2 years ago
On Fri, 6 Feb 2004, Julia Engelmann wrote: > > Hi, > I am comparing different normalizations of Affymetrix microarrays: > 1. expresso with rma, vsn, pmonly and medianpolish > 2. calling rma and vsn with: eset=rma(Data); Set=vsn(eset); > I know that medianpolish does log2 transformation and vsn does log > transformation (base e), but when I compare the results, the difference > between the two datasets is huge- not a matter of log-base. > Shouldn't the results be very similar? > Julia, vsn doesn't use log (base) but a different trasnformation that stabilizes the variance, I have found the results to be quite different too, they are meant to be especially if there is a strong dependency between variance and expression levels. Best Susan Holmes Associate Professor Statistics Stanford
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@anja-von-heydebreck-625
Last seen 10.2 years ago
Hi Julia, >Hi, >I am comparing different normalizations of Affymetrix microarrays: >1. expresso with rma, vsn, pmonly and medianpolish >2. calling rma and vsn with: eset=rma(Data); Set=vsn(eset); > rma gives you expression levels on the log scale, whereas vsn expects input values on the raw scale. So it doesn't make sense to call vsn after rma like in 2. (apart from the question why you should normalize your data twice - I guess in 1. you used rma only for background correction; but calling rma like in 2. involves also quantile normalization and medianpolish). >I know that medianpolish does log2 transformation and vsn does log >transformation (base e), but when I compare the results, the difference >between the two datasets is huge- not a matter of log-base. >Shouldn't the results be very similar? > >Any help would be much appreciated. > >Thanks, >Julia > > Best, Anja
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On Sat, 7 Feb 2004, Anja von Heydebreck wrote: > Hi Julia, > > >Hi, > >I am comparing different normalizations of Affymetrix microarrays: > >1. expresso with rma, vsn, pmonly and medianpolish > >2. calling rma and vsn with: eset=rma(Data); Set=vsn(eset); > > > rma gives you expression levels on the log scale, whereas > vsn expects input values on the raw scale. So it doesn't make sense > to call vsn after rma like in 2. (apart from the question why you > should normalize your data twice - I guess in 1. you used rma only for > background correction; but calling rma like in 2. involves also > quantile normalization and medianpolish). of cource you could do: exprs(eset) <- 2^exprs(eset) eset = vsn(eset) exprs(eset) <- log2(exprs(eset)) this should make things look more alike. > > >I know that medianpolish does log2 transformation and vsn does log > >transformation (base e), but when I compare the results, the difference > >between the two datasets is huge- not a matter of log-base. > >Shouldn't the results be very similar? > > > >Any help would be much appreciated. > > > >Thanks, > >Julia > > > > > Best, > Anja > > _______________________________________________ > Bioconductor mailing list > Bioconductor@stat.math.ethz.ch > https://www.stat.math.ethz.ch/mailman/listinfo/bioconductor >
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Rafael A. Irizarry wrote: >On Sat, 7 Feb 2004, Anja von Heydebreck wrote: > > >>Hi Julia, >> >> >>>Hi, >>>I am comparing different normalizations of Affymetrix microarrays: >>>1. expresso with rma, vsn, pmonly and medianpolish >>>2. calling rma and vsn with: eset=rma(Data); Set=vsn(eset); >>> >>> >>> >>rma gives you expression levels on the log scale, whereas >>vsn expects input values on the raw scale. So it doesn't make sense >>to call vsn after rma like in 2. (apart from the question why you >>should normalize your data twice - I guess in 1. you used rma only for >>background correction; but calling rma like in 2. involves also >>quantile normalization and medianpolish). >> >> > >of cource you could do: > >exprs(eset) <- 2^exprs(eset) >eset = vsn(eset) > But the output of vsn is - at least approximately - on the natural log scale. >exprs(eset) <- log2(exprs(eset)) > Therefore, to obtain log2-values, I would rather do exprs(eset) <- log2(exp(exprs(eset))), or exprs(eset) <- log2(exp(1))*exprs(eset) Best, Anja -- Dr. Anja von Heydebreck Max Planck Institute for Molecular Genetics Dept. Computational Molecular Biology Ihnestr. 73 14195 Berlin, Germany heydebre@molgen.mpg.de phone: +49-30-8413-1168 http://www.molgen.mpg.de/~heydebre fax: +49-30-8413-1152
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Dear all, I am working in R a program for cross validation. I have a vector of 20 observations and I want to find all the possible combinations (20,4) and store them as data. I wonder if someone can help me. Thanks in advance. Makis ---------------------- E Motakis, Mathematics E.Motakis@bristol.ac.uk
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wrong place for this question, should go to r-help@r-project.org anyway, i am not aware any R-function that gives all subsets of size k. i will use matlab to create it and input it back to R if you don't want to stick with matlab for the rest. if you have some time to spare, you can create your own function by implement the algorithms that generates all subsets of size k. a good source for such algorithms is http://www.cs.sunysb.edu/~algorith/ good luck Kenny Kenny Ye Assistant Professor Department of Applied Math and Statistics SUNY at Stony Brook Stony Brook, New York 11794-3600 Phone (631)632-9344 Fax (631)632-8490 On Mon, 9 Feb 2004, E Motakis, Mathematics wrote: > Dear all, > > I am working in R a program for cross validation. I have a vector of 20 > observations and I want to find all the possible combinations (20,4) and > store them as data. I wonder if someone can help me. > > Thanks in advance. > Makis > > ---------------------- > E Motakis, Mathematics > E.Motakis@bristol.ac.uk > > _______________________________________________ > Bioconductor mailing list > Bioconductor@stat.math.ethz.ch > https://www.stat.math.ethz.ch/mailman/listinfo/bioconductor >
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Hi E, have a look at the function "nchoosek" from package vsn. Best wishes Wolfgang E Motakis, Mathematics wrote: > Dear all, > > I am working in R a program for cross validation. I have a vector of 20 > observations and I want to find all the possible combinations (20,4) and > store them as data. I wonder if someone can help me. > > Thanks in advance. > Makis > > ---------------------- > E Motakis, Mathematics > E.Motakis@bristol.ac.uk > > _______________________________________________ > Bioconductor mailing list > Bioconductor@stat.math.ethz.ch > https://www.stat.math.ethz.ch/mailman/listinfo/bioconductor -- ------------------------------------- Wolfgang Huber Division of Molecular Genome Analysis German Cancer Research Center Heidelberg, Germany Phone: +49 6221 424709 Fax: +49 6221 42524709 Http: www.dkfz.de/abt0840/whuber
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