[BioC] limma and paired data
Naomi Altman
naomi at stat.psu.edu
Mon Jun 7 02:03:58 CEST 2004
The simplest way to handle this is probably to take the difference of the
pairs and feed that into limma as a 1-sample t.
--Naomi
At 12:22 PM 4/20/2004 +1000, Gordon Smyth wrote:
>At 03:09 AM 20/04/2004, Danielle Fletcher wrote:
>>Hi,
>>
>>I am using limma to analyse a 2-colour microarray experiment. There are 2
>>treatments and 4 replicates in each of these groups. Each replicate is
>>paired to a replicate in the otehr treatment group. Each sample was
>>hybridised with a reference, so 8 slides in total.
>>
>>The targets file looks like this (hopefuly that will make it clearer):
>>SlideNumber Name FileName Cy3 Cy5
>>1 1M 1.gpr monolayer ref
>>2 1P 5b.gpr pellet ref
>>3 2M 2.gpr monolayer ref
>>4 2P 7.gpr pellet ref
>>5 3M 3.gpr monolayer ref
>>6 3P 6.gpr pellet ref
>>7 4M B.gpr monolayer ref
>>8 4P A.gpr pellet ref
>>
>>Initially my design matrix looked like this:
>>
>> Sample-Ref Monolayer-Pellet
>>1M 1 0
>>1P 1 1
>>2M 1 0
>>2P 1 1
>>3M 1 0
>>3P 1 1
>>4M 1 0
>>4P 1 1
>>
>>but thinking about it again, i don't think this takes into account the
>>paired nature of the data. I am sure that the answer is probably a simple one,
>
>There is no simple answer. There was a big discussion about this in
>Bioconductor very recently, please look at the list archives.
>
>In the very latest versions of limma, there is a new argument 'block' in
>duplicateCorrelation() and lmFit() to handle a blocking structure like you
>describe. This feature is however very lightly documented so far and so is
>offered on a user beware basis. In your case, block=c(1,1,2,2,3,3,4,4).
>
>Gordon
>
>> but I am not sure what the best solution is. I would appreciate any
>> advice.
>>
>>Thanks in advance
>>
>>Danielle
>
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Naomi S. Altman 814-865-3791 (voice)
Associate Professor
Bioinformatics Consulting Center
Dept. of Statistics 814-863-7114 (fax)
Penn State University 814-865-1348 (Statistics)
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