Dan Eilat wrote:
>> Looking urgently for a procedure to make a good mRNA from rat kidney
> glomeruli (in order to make cDNA library).
> Thanks., Dan Eilat (my e.mail is: eilat at md2.huji.ac.il)
Hi Eilat!
After having isolated the glomeruli, homogenize them in TRI-reagent (or
RNA-Clean, Trizol....) according to the protocol supplied with the
reagent. Also the RNA, DNA and protein isolation will follow the given
protocol.
This works very well in our labs.
We also isolated glomerular mesangial cells according to:
Lovett D. H., Ryan J. L., Sterzel R. B. (1983)A thymocyte-activating
factor derived from glomerular mesangial cells. J. Immunol. 130,
1796-1801
and used them as primary cultures to passage 25. For RNA preparation
from this cells we also used RNA-Clean or TRI-reagent.
hope this helps....
bye
Heinz
--
Heinz-Juergen Schaefers
University of Erlangen-Nuernberg
Dept. of Medicine IV
Nephrology Research Lab
h-j.schaefers at popHost.com
