tRNA elution buffer?

Lyle Najita ijiwaru at nyc.pipeline.com
Wed Jul 19 19:00:54 EST 1995


In bionet.general jammi at SELWAY.UMT.EDU (Narasimham Jammi) said: 
 
 
>Hi, 
> 
>I have had problems in eluting out tRNA out of denaturing PAGE. Could
anybody  
>suggest a good elution buffer. Thank you. 
> 
>jammi 
> 
>-------------------------------------------------------------------------------
 
 
 
Jammi, 
 
this is a receipe that works for me: 
 
0.5M    NH4OAc 
10mM  MgCl2  
0.1%    SDS 
0.1mM  EDTA 
 
We usually elute at 4 degrees C, twice overnight and precipitate the RNA
with glycogen (10-30ug) and 3 volumes of ethanol.  Recovery varies with the
percentage of the gel and the size of the RNA.  For T7 transcripts of about
300nt isolated on 5% acrylamide (19:1 acrylamide/bisacrylamide)/ 7M urea
gels I would estimate about 70-80% recovery of counts.  Hope this helps. 
 
Lyle Najita 
Rockefeller University 



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