Brain Total RNA extraction

Bob Hartley r.hartley at bio.gla.ac.uk
Fri Oct 12 04:45:11 EST 2001


In article <3bc6929f.745958 at news.task.gda.pl>,
 wasowicz at uwm.edu.pl (Krzysztof W. Wasowicz) wrote:

> On 11 Oct 2001 18:30:59 -0000, Adam <adam.kuhl at usm.edu> wrote:
> 
> >I am trying to extract total RNA from fish brain tissue using Trizol
> >reagent, but am getting very low A260/A28.  I have seen that this could be
> >due to high lipid content.  Can someone give me a good method of solving
> >this problem?
> >
> 
> >Thanks,
> 
> >Adam
> >
> >
> ><http://www.biowww.net/forum/read.php?f=1&i=4121&t=4121>
> >
> Hi,
> I have noticed that I am getting better results with an original
> method of Chomczynski & Sacchi (Anal Bioch - you can find reference
> from Medline probably - if not, write me). 

You might better using a GITC lysis then a seperate acid phenol 
chloroform extraction.

you might also consider using a tissue disruptor likre the bio101 
machine (also sold by Hybaid) OR rather than buying the machine you 
could buy the beads and use a whirlymixer.


also you could try freezing the lysis solution to -70/80 AND RAPIDLY 
THAWING IT IN A WATER bath (damn caps lock). :-)

you can make your own -70 desktop freezer for 10pounds ~16euros

the detais are available inthe archive. see posting by me Robert Hartley
rh at mblab.gla.ac.uk
  
cheers
Bob; Sunny Scotland



-- 
Robert Hartley
Spinal Cord Research Group
University of Glasgow. Glasgow G12 8QQ
mail: r.hartley at bio.gla.ac.ukWeb : 
http://www.gla.ac.uk/ibls/NBS/spinal.html




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