RNAse showing up in gel after plasmid miniprep
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.
On 17 Dec 1996, Marianne Leverone ,
> 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"
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
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