RNAse showing up in gel after plasmid miniprep

Ken Howe howe at DARWIN.UCSC.EDU
Tue Dec 17 15:42:13 EST 1996


To remove RNase from our DNA preps (after RNase treatment, for example), 
we treat with proteinase K (DNase-free) followed by one or more
extractions (ie., with phenol then phenol:chl:IAA, etc).  Precipitation
does not remove or inactivate RNase.

Hope this helps.

Ken Howe

On 17 Dec 1996, Marianne Leverone ,
BIO wrote:

> Date: 17 Dec 1996 08:02:26 -0800
> From: "Marianne Leverone , BIO" <leverone at chuma.cas.usf.edu>
> To: methods at net.bio.net
> Subject: RNAse showing up in gel after plasmid miniprep
> 
> Hi Group!  When we run plasmid minipreps (alkaline lysis), we add RNAse
> after the white protein/DNA junk has been pelleted and removed from the
> neutralization step.  After a 30 min incubation, the plasmid is then ppt
> with isopropanol and 70% etOH washed and dried and resuspended in water.
> Then a gel is run to check for complete RNA removal.  Some of our labgroup
> people are having trouble getting rid of the RNase becasue it shows up as
> a band (around the 0.5 kb marker) and we have now run enough controls
> and other tests and have determined that the band is RNase.  ANyone else
> have this problem and how did you solve it?  Thanks.  Marianne
> 
> 
> 


               "You know, it sure would be amino world without RNA"
                     _______________________________________

                                  Kenneth Howe
                     Center for the Molecular Biology of RNA
                             Department of MCD Biology
                             University of California
                              Santa Cruz, CA  95064
                     _______________________________________

                          e-mail: howe at darwin.ucsc.edu
               http://www-biology.ucsc.edu/people/areslab/ken.html




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