Question about Formaldehyde gels and Northern Blots

[Susanne Rohrer]

John Thompson wrote:

> >3) Could the RNA be degrading in the Formaldehyde gel?
>
> If you are just using the RNA for hybridizations, try dissolving the
> RNA in 0.02% DEPC. This will inactivate any potential RNAse that may
> have carried through with the prep.

Our results changed dramatically when we started using Formamide for
dissolving the RNA preps. unfortunately you can only use it for Northerns,
for Enzymatic reactions you have to keep using water.

> >2) Are we using the correct transfer buffer for Nylon membranes?  We are
> >doing a transfer overnigth with 10X SSC.

Maybe you should try vacuum transfer. it works really well.


Susanne