[BioC] extracting significant genes using limma
James W. MacDonald
jmacdon at med.umich.edu
Tue Mar 14 15:41:58 CET 2006
Naomi Altman wrote:
> Dear Assa,
> This one will need to go to the list. I only use topTable.
>
> --Naomi
>
> p.s. Please post all questions directly to the listserv. That way
> you can get more help than any one person can give, and no-one is
> over-burdened.
>
> At 03:29 AM 3/14/2006, you wrote:
>
>>Hi,
>>
>>I was wondering if I can use decideTests()-function in my tests,
>>even if i didn't use any contrast matrix.
Yes you can. However, depending on how you set up your design matrix you
may or may not get the comparisons you are looking for.
>>
>>I didn't get the difference between decideTests() and classifyTests().
decideTests() is a wrapper function that calls classifyTestsP() or
classifyTestsF() if you select method = "heirarchical" or "nestedF",
respectively. decideTests() also uses different default p-value cutoffs
and multiplicity corrections. Looking at the code for decideTests() may
help you understand the differences.
HTH,
Jim
>>
>>I would be happy for any help you can give me.
>>
>>THX
>>
>>Assa
>>
>>On 3/13/06, Naomi Altman
>><<mailto:naomi at stat.psu.edu>naomi at stat.psu.edu> wrote:
>>Since you used "adjust=fdr", the p-value column of the TopTable are
>>the "adjusted p-values" after fdr (which I think of as q-values).
>>
>>You can either pick some q-value you want to use to select the
>>significantly differentially expressing genes, or you can pick some
>>number of genes, and report the q-value of the least significant of these.
>>
>>--Naomi
>>
>>At 10:17 AM 3/13/2006, Assa Yeroslaviz wrote:
>>
>>>Hi,
>>>
>>>I know this theme is an old one, but I look all over the archives and didn't
>>>find any help regarding this subject.
>>>Using Affymetrix chips I compared two groups (Control vs compound) with the
>>>limma procedure.
>>>I made an affybatch Object using ReadAffy(), normalised the data with the
>>>RMA algorithm and fitted a linear model with lmFit.
>>>
>>>
>>>>affy <- ReadAffy(filenames=vec)
>>>>eset <- rma(affy)
>>>>design <- cbind(Control=1,AE0627vsCT=c(rep(0,6),rep(1,4)))
>>>
>>>my design matrix looks like that (I have 6 control and 4 treated arrays):
>>>
>>>>design
>>>
>>> Control AE143vsCT
>>> [1,] 1 0
>>> [2,] 1 0
>>> [3,] 1 0
>>> [4,] 1 0
>>> [5,] 1 0
>>> [6,] 1 0
>>> [7,] 1 1
>>> [8,] 1 1
>>> [9,] 1 1
>>>[10,] 1 1
>>>
>>>so I don't need any contrast matrix.
>>>The list is 22,810 genes long. But not all of them can be significant. I
>>>hope!!!
>>>
>>>I sorted the genes with:
>>>
>>>>sig_table <- topTable(fit_e, coef=2, number=6000, adjust="fdr", sort.by=
>>>
>>>"P")
>>>
>>>I've chosen 6000 as an arbitrary value, but I still don't know how many
>>>genes are siginificant.
>>>
>>>My question(s) is(are):
>>>
>>>1. How do I find out how many genes are significantly differentially
>>>expressed using a define p-value or FDR?
>>> Can I use here the decideTests() function although I don't have any
>>>contrasts?
>>>
>>>2. In SAM one can look for the false discovery rates using the different
>>>delta-values.
>>> Is it possible to set a fixed FDR-Value in Limma?
>>> Where Do I find the FDR rates of my significant genes?
>>>
>>>3. Is there a possibility (like in SAM) to show the results in a graphic (
>>>scatter plot etc.)?
>>>
>>>Every comment and suggestion would be appreciated!
>>>
>>>THX
>>>
>>>Assa
>>>
>>>--
>>>Assa Yeroslaviz
>>>Loetzener Str. 15
>>>51373 Leverkusen
>>>
>>> [[alternative HTML version deleted]]
>>>
>>>_______________________________________________
>>>Bioconductor mailing list
>>><mailto:Bioconductor at stat.math.ethz.ch>Bioconductor at stat.math.ethz.ch
>>>https://stat.ethz.ch/mailman/listinfo/bioconductor
>>
>>Naomi S. Altman 814-865-3791 (voice)
>>Associate Professor
>>Dept. of Statistics 814-863-7114 (fax)
>>Penn State University 814-865-1348 (Statistics)
>>University Park, PA 16802-2111
>>
>>
>>
>>
>>--
>>Assa Yeroslaviz
>>Loetzener Str. 15
>>51373 Leverkusen
>
>
> Naomi S. Altman 814-865-3791 (voice)
> Associate Professor
> Dept. of Statistics 814-863-7114 (fax)
> Penn State University 814-865-1348 (Statistics)
> University Park, PA 16802-2111
>
> [[alternative HTML version deleted]]
>
> _______________________________________________
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> Bioconductor at stat.math.ethz.ch
> https://stat.ethz.ch/mailman/listinfo/bioconductor
--
James W. MacDonald, M.S.
Biostatistician
Affymetrix and cDNA Microarray Core
University of Michigan Cancer Center
1500 E. Medical Center Drive
7410 CCGC
Ann Arbor MI 48109
734-647-5623
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