RE RNA from fat

John Brennand, Tel 4639/5739/6636 John.J.C.Brennand at APVXC1.Pharmaceuticals.ZENECA.tmailuk.sprint.com
Mon Aug 14 09:06:04 EST 1995


We have been making RNA from mouse adipose tissue recently for PCR & 
northerns.  We remove adipose tissue from the animals and store it at -80.  We 
then process 0.5 gm following the Trizol (Life Technologies) protocol.  There 
is a lipid interface that needs avoiding when removing the aqueous phase.  I 
never bother re-extracting the organic phase although that should increase 
yields somewhat.  After precipitation and re-suspesion in water, this 
typically yields about 200 ug of total RNA.  We then purify this on OligotexdT 
(Qiagen) and routinely get about 10 ug of polyA+ material. Both total and 
purified material look excellent on a gel and blot well with control probes.  
We have also succesfully PCRd a number of full length cDNA products from this.

Hope this helps, good luck.



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