northern blot help needed

z z-suldan at ski.mskcc.no.spam.org
Fri Nov 7 07:35:46 EST 1997


In article <63ss6h$jbo at whitbeck.ncl.ac.uk>, laura gray
<laura.gray at ncl.ac.uk> wrote:

> I am using Capillary blot method for northern blotting as in Maniatis. 
> Recently about 50% of the RNA has not been transferring to the membrane ( 
> Genescreen plus). i have tried new batches of buffer and agarose to no 
> avail. Can anyone help?

Just some random thoughts that helped me out tremendously.

Using the cap action transfer, put the gel on upside down so the bottom of
the gel is what is in contact with the membrane. Outline the gel with thin
strips of parafilm so the wicks don't short circuit. And don't use too
much weight. I use two glass plates (~ 5" x 8" ??) and that's it.

My last Northern transfered completely in under 6 hrs.

Good luck.

Zal



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