Warning about DNaseI treament of RNA

ELADER at AMBION.COM ELADER at AMBION.COM
Sun Sep 20 11:12:18 EST 1998


In article <pnorton-ya023080001809981244090001 at tjnews.tju.edu>,
  pnorton at lac.jci.tju.edu (Pamela Norton) wrote:
> In article <6tsdut$c32$1 at nnrp1.dejanews.com>, ELADER at AMBION.COM wrote:
>

>
>      Do you know at what temperature the problem starts?
> We have seen this phenomenon manifested in two ways. First, when we run

Northerns of RNA that have mg++ in it. We add loading dye, heat to 65 degrees
for 10 minutes, and run the gel, blot and probe. We get much lower signal if
the RNA had more than 3 mM MgCl2 in the sample. To insure against the
problem, add enough EDTA to the loading dye.

The second place we see it is when we  DNase an RNA probe prior to running on
a page urea gel. Again, EDTA will cure the ills.

So......65-68 degrees with over 3 mM MgCl2

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