Vacuum Blotter - RNA transfer

VASANTHARAJAN JANAKIRAMAN vasant at cc.UManitoba.CA
Sun Apr 9 11:05:40 EST 2000


After fiddling with the set up-porous screen with the gaskets, last time
after 3 hrs. (in 10xssc), all my wheat RNA got transferred into the hybond
N+ membrane.
vasanth

 On Sun, 9 Apr 2000, Mano wrote:

> Date: Sun, 09 Apr 2000 07:34:06 GMT
> From: Mano <mdep at musica.mcgill.ca>
> To: methods at hgmp.mrc.ac.uk
> Subject: Re: Vacuum Blotter - RNA transfer
> 
> By the way, how long did you transfer using the vacuum? I ask because I use vacuum
> blotter.  Thanks
> 
> 
> Vellanoweth's Lab wrote:
> 
> > We've recently discarded the vacuum blotter for northern transfer.  We are now
> > back to the tried and true papertowel method for blotting which works
> > beautifully and is actually much easier, if a little longer in time.  We found
> > over half our sample was still in the gel with the vacuum blotter and we had
> > bad hybridisation success, now we are hybridising fine, with all the sample
> > there.
> >
> > Janel Wheeler
> > Vellanoweth Laboratory
> > Cal State Univ., Los Angeles
> > jwheele at calstatela.edu
> >
> > Susanne Rohrer wrote:
> >
> > > VASANTHARAJAN JANAKIRAMAN wrote:
> > >
> > > >
> > >
> > > > Even after 2 hrs of transfer at 55mbar, my RNA (Wheat RNA) was still in
> > > > the gel.
> > >
> > > We see the same when using bacterial total RNA . There are lots left in the
> > > gel, but what is transferred is still enough for detection - have you tried
> > > hybridization and detection on it?
> > >
> > > Susanne
> 
> 

---




More information about the Methods mailing list