RNA Pellet viscosity (Pectic acid?)
jorgen.borrebak at basalmed.uio.no
Thu Jun 3 12:48:46 EST 1999
I don't know about pectin, but I've read and heard that polysaccharides,
or even proteoglycans, might be a considerable contaminant in
When I isolate RNA from white blood cells (wich are rich in RNAses and
other substances like lipids, and relatively lean on the RNA side) I use
guanidine thiocyanate lysis, one acid phenol extraction and one
This removes a lot of "goo".
After dissolving in water I go on with Dynal's oligo dT magnetic beads.
I get some precipitation of this "substance X", but not too much to
interfere with the beads.
The mRNA exctracted should be perfect for RT-PCR, only I havn't tried it myself.
In article <37497E28.121B5704 at sph.unc.edu>, John Scott Meschke
<jmeschke at sph.unc.edu> wrote:
> In my earlier message I was asking about two phases (aqueous and
> gelatinous) produced from resuspension of RNA pellet after alcohol
> precipitation. A friend has suggested that the problem could be pectin
> (or pectic acid). The environmental sample that I was trying to recover
> spiked viral RNA from was a chloroform extract of tomato sauce. Has
> anyone seen this problem before (with RNA extracts from plants,
> maybe?)? Is the pectic acid theory sound? What steps would I take to
> remove the pectic acid from the RNA or vice versa? Any other ideas?
> Any help will be much appreciated.
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