[BioC] experiment-design-advice-request
Naomi Altman
naomi at stat.psu.edu
Sun May 1 17:25:36 CEST 2005
Design 1 is better than 2 but is unbalanced for the dye effect. If there
could be a dye by gene interaction you may have some difficulties in
interpreting the design.
Design 2 is unbalanced for the main effects. You would be better to
complete the loop, with 4 hybridizations.
I am not sure what you mean by "dyeswap and 2 replicates". Are you using 6
or 3 arrays, or is this the base design which you intend to replicate? If
you plan to have 8 or 12 arrays, you have many more choices for the design.
Finally, technical replication is not that informative. If at all
possible, each hybridization should be done with an independent biological
sample. Technical replication is done mainly if the cost of creating an
independent biological sample is too high. (Technical replication is also
done due to the misconception that it is necessary for dye-swapping.)
Just a note - statisticians consider the control to be a condition, so you
actually have 2 conditions.
--Naomi
At 02:49 PM 4/30/2005, you wrote:
>hi friends
>we are designing a microarray experiment, where there are two different
>mouse strains (A,B)...
>and one condition (Peptide - P and Saline - S). we expect the mouse
>strains to express differentially even under normal (saline)
>conditions...so we did not want to go for pooling the controls, to have a
>common - pooled control... under this scenario...
>
>which of the following designs would you suggest....(we have planned to
>use dyeswap and 2 replicates for each hybrisation)....
>
>1:
>
>AP---BP
>| \ / |
>| / \ |
>AS---BS
>
>(all pairs - with 6 hybridisations)
> OR
>
>2:
>
>AP
>|
>AS---BS
> |
> BP
>
>(with just 3 hybridisations, so that we could deduce AP-BP,using AP-AS-BS
>and rest like that...)
>
>i request your valuable advice in this regard....
>thanks
>
>vijay
>graduate student
>department of biological sciences
>University of Southern Mississippi
>MS
>
>
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>
>
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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)
University Park, PA 16802-2111
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