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Whole Genome Analysis of small Antarctic tissue samples

WGA small sample size

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#1 AFGPhunter

AFGPhunter

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Posted 19 April 2012 - 03:59 PM

Hello everyone,

I am trying to find a specific gene motif in an Antarctic animal which I have reason to believe jumped species in a horizontal gene transfer event. The main problem is my limitation on the sample size and techniques. This work has to be done from juvenile eggs of the species as the adults live parasitic on fish and that is the source of the gene I am after. So I need to exclude any fish contamination and the only way to do so is working with eggs.
And I have probably cell clusters for each egg of maybe a few 100 cells. I can pool around 50 of them but they weigh each around 1ug tops, and most of it is water and probably nutritional deposits from the adults.

I was wondering whether anyone has worked with a few cells from wild animals for a WGA yet and neede to boost the gDNA or library?
I also wonder who might be willing to provide me with some hands on experiences on the use of gDNA amplification tools like the GenomiPhi kit?
Anyone who worked on samples of marine species with mucus and saccharids as trouble making molecules for WGA?
Has anyone indications to clonality issues as encountered by ancient DNA teams when using DNA amplifications for WGAs?

I would appreciate very much indeed any comments and helpful hints or pointers to groups who might be willing to help!





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