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YUK FAI LEUNG
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140
@yuk-fai-leung-605
Last seen 10.3 years ago
Hi there,
I am designing a pilot microarray study on embryoic developmental
mutant
using affy platform. The comparison itself is very simple, the mutant
vs
normal at one time point. Due to various reasons (mostly funding and
limited amount of tissue), I can't start with the "ideal" approach in
which each sample is hybridized to an individual chip.
Since I can easily rear a lot of animals, it seems that pooling is the
only choice for the pilot study. However I am not sure what is the
best
way to allocate the pooled samples to each chip. For example if I want
to do 3 array replicates each for the mutant and control. Is it better
to pool enough samples for 3 arrays and then separate the pooled
sample
in 3 portions for hybridization or just pool different individual
samples for different replicates?
It seems to me that the first way is like getting a group expression
average with accessment of technical variation, while the second
approach can also provide some sort of evalution of biological
variation, abeit an averaged one by the pooling. I suspect the latter
approach is better, and would love to know the suggestions from you.
Thanks!
Fai
--
Yuk Fai Leung
Department of Molecular and Cellular Biology
Harvard University
BL 2079, 16 Divinity Avenue
Cambridge, MA 02138
Tel: 617-495-2599
Fax: 617-496-3321
email: yfleung@mcb.harvard.edu; yfleung@genomicshome.com
URL: http://genomicshome.com