Warning about DNaseI treament of RNA

Pamela Norton pnorton at lac.jci.tju.edu
Fri Sep 18 12:44:09 EST 1998


In article <6tsdut$c32$1 at nnrp1.dejanews.com>, ELADER at AMBION.COM wrote:

> Some recent posts about DNase I treatment prompt me to warn those of you who
> are doing this about a general phenomenon we have observed at Ambion.
> 
> If you adjust the buffer conditions of your RNA samples to above a couple of
> mM Mg++ and you then heat the sample later (to kill the DNase for example)
> the RNA may be degraded in the heating process. This holds true for total RNA
> and for in vitro transcribed RNA.
> 
> If you must add Mg++ to any RNA sample over 3 mM, before you heat you must add
> EDTA to a level above the level of the Mg.
> 
> Eric
> 

Eric,

     Do you know at what temperature the problem starts? 

     Pam Norton

-- 
Pamela A. Norton, Ph.D.          Associate Professor of Medicine
Thomas Jefferson University
Philadelphia, PA 19107           p_norton at lac.jci.tju.edu



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