Hi everybody. I am a little bit confuse regarding to pool the sample or not to pool the sample regarding the human proteome sample I have. What should I do for 2DE analysis? Should I pool e.g. 6 samples and run triplicate or run 6 2DE gels?
I would be glad if you could give me some reference.
Merci,Kambiz
To understand whether or not pooling is a good idea, it would help if you stated the objectives of the experiment. Pooling can be useful if you're looking for comprehensive fingerprint, but it masks the differences between individuals in a population.
In my line of work (developing assays); I generally avoid pooled samples in most experiments as they tend to hide interference resulting from non-analyte species.
Hi Andrew. thank you for your comment. It is a pilot study for a bigger plan for developing a diagnostic test through proteomics technology. I have plasma and seminal plasma sample. I have extracted out the proteome and want to do 2DE in first step to have some candidates.
Merci, Kambiz
In which case, I would only use pooled serum as a calibrator and/or controls. As I stated above, pooled samples will mask any interference in your assay.
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