RNAse showing up in gel after plasmid miniprep

George Rutherford gruther at bilbo.bio.purdue.edu
Mon Dec 23 12:39:08 EST 1996

In article <199612200015.RAA15827 at NMSU.Edu>, hroychow at NMSU.EDU (Hiranya
Roychowdhury) wrote:

> At 04:50 PM 12/18/96 GMT, Paul N Hengen wrote:
> >Marianne Leverone ", BIO (leverone at CHUMA.CAS.USF.EDU) wrote:
> >
> >> Berrnard M:  We are using <1 ug/ml.  Only one person in our lab is having
> >> this trouble which is wierd.  We can't figure out why he is having such
> >> trouble!  He is now trying a phenol:chloroform then chloroform extraction
> >> to see if he can get rid of it.  this would be a pain because these are
> >> minipreps, and the templates (a lot of them) are being used for DNA
> >> sequencing (for mutation screening).  Marianne
> >
> >Add MORE RNase. What you are seeing on the gel is the RNA and not RNase!
> >
> >--
> >* Paul N. Hengen, Ph.D.                          
> >* National Cancer Institute                       |Internet:
pnh at ncifcrf.gov |*
> >************************************
> >
> >
> But RNA runs well below the 0.5kb marker band, doesn't it? If I recall
right, the band Marianne originally wrote about is at 0.5kb. Although, I
have to admit, I have never seen RNase band in any of my gels (and I have
run thousands by now)!!

Ladies and gentlemen,

Do the experiment. Equivalent amount of RNase with no DNA. Miniprep DNA
with no RNase. Personally, I think the 0.5kb stuff is almost certainly
RNA, but one nevers knows, I suppose.


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