granulosa cell RNA

r.l.bates bch140 at sysa.abdn.ac.uk
Mon Apr 27 05:53:40 EST 1998


I'm trying to extract RNA from human granulosa cells collected during oocyte
aspiration for IVF. I intend to make cDNA.

At the moment I am using the Chomaski and Sacchi method after extracting the
granulosa cells. 

The first time I tried extracting RNA I cultured the cells first and I got RNA,
since then I have just been collecting the cells and going straight to RNA
extraction and getting nowhere.

Do I need to culture them first or do I need to get hold of the oocyte aspirates
more quickly and put them on ice? I am still using the same extraction procedure
for the granulosa cells as for culturing them which keeps them at room
temperature. 

Any suggestions would be appreciated.

Thanks

R.L.Bates
Depts of Obstetrics and Gynaecology, and Molecular and Cell Biology,
University of Aberdeen,
Scotland, UK.



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