[BioC] LIMMA design question

Wei Shi shi at wehi.EDU.AU
Fri Apr 8 23:08:24 CEST 2011


Hi Prasad:

	I think you can treat patients from the same disease as replicates because they are actually biological replicates but not technical replicates. You contrasts for comparing diseases to healthy also look OK ...

Cheers,
Wei

On Apr 9, 2011, at 3:59 AM, Prasad Siddavatam wrote:

> Hello Group,
> 
> I am analyzing individual patient arrays and need some help on the following 
> questions.  
> 
> 1. If I have a dataset with four types of patients (disease1, disease2,
> disease3, healthy) and want to get the differential genes between disease 
> and healthy. I can get the contrasts between healthy and disease 
> comparisons using contrasts in LIMMA. But here each disease has 2-3 
> arrays from different patient blood samples. These are not true
> replicates, per se but thats all we have. My question is how do we 
> treat them? Can I consider them as replicates? (I understand LIMMA will 
> combine the replicate data.)
> here is the design........
> 1. disease1 patient1
> 2. disease1 patient2
> 3. disease1 patient3
> 4. disease2 patient4
> 5. disease2 patient5
> 6. disease3 patient6
> 7. disease3 patient7
> 8. disease3 patient8
> 9. healthy patient1
> 10. healthy patient2
> 11. healthy patient4
> 
> 2. In the above scenario, If I want differentially expressed genes in 
> disease1 and disease2 combined w.r.t to healthy, is the following is 
> correct way in LIMMA?
> makeContrats(dis1&dis2 <- "(disease1 + disease2) /2 - healthy", 
>                   levels = design);
> 
> If I want differentially expressed genes in all three diseases combined
> w.r.t to healthy 
> makeContrats(dis <- "(disease1 + disease2 + disease3) /3 - healthy", 
>              levels = design);
> 
> please let me know is it the right approach.
> 
> Greatly greatly appreciate your help,
> 
> Sincerely
> 
> Prasad
> 
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