Please, help with RNA extraction.

Dr Richard Sallie sallier at cygnus.uwa.edu.au
Sat Jun 15 20:47:39 EST 1996


This is because of the high lipid content of brain and liver.  It is
important to chill the tissue-extraction fluid homogenate before
centrifugation (or to do it in a cold centrifuge 4oC).  Occasionally
increasing the amout of chloroform in the mix and back extracting the
organic phase with Na Acetate solution and repeated chloroform (Chisam)
extraction helps here.  The degree of difficulty of liver extractions
depends on the amount of fat in the liver, which for some animals (and
experiments) is diet (and hibernation cycle) dependent.  The usual
disclaimers.
Good luck !
Richard Sallie MD
In article <melia.73.0 at ar.vhebron.es>, melia at ar.vhebron.es wrote:

> I have problems when I try to isolate total RNA from mouse liver or brain by 
> Chomczynsky method, because the two phases I should obtain after adding 
> phenol and chloroform doesn't separate during centrifugation. This 
> difficults the extraction and makes the efficiency to be very low.Does 
> anybody know why this happens and how to avoid? Is this method good enough  
> to extract RNA from those tissues?
> 
> I would appreciate your comments a lot.
> Thank you,
> 
> Maria
> 
> melia at ar.vhebron.es



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