[BioC] Normalizing by Within Condition - a bad idea

Kasper Daniel Hansen khansen at stat.Berkeley.EDU
Tue Feb 28 07:25:10 CET 2006


Hi Naomi

I would tend to agree with you. However, there are numerous (ok,  
some) applications where the (just assume simple treatment/control  
setup) treatment is expect to be massively different than the  
control. Say eg. if treatment is cyclohexamid. As basically all  
normalization techniques were developed for the "largely all genes  
unchanged" situation, it is not clear how one would approach a  
massive global change.

However, come to think of it, the examples I have in mind are not  
exactly standard expression studies. In fact they are not even  
expression studies.

/Kasper

On Feb 27, 2006, at 7:30 PM, Naomi Altman wrote:

> The question of whether normalizing by condition is reasonable
> frequently comes up on this list.  For a recent talk, I took the
> first 2 conditions (experiments) of the Affymetrix Latin Square
> spikein experiment and normalized by RMA all together and within
> treatment.  The experiment uses identical RNA except for 42 spike-ins
> which are varied by condition.  There are 3 replicates, making 6  
> arrays in all.
>
> The number of "significant" genes at p<.01 using limma with eBayes
> (but no multiple comparison adjustment) is 144 (capturing most but
> not all of the spike-ins) when the arrays are normalized together and
> 14374 when they are normalized by experiment.
>
>
> 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
>
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