Re-using Northern Blots?

Melissa Melan MMELAN at LUCY.WELLESLEY.EDU
Tue Feb 2 12:01:00 EST 1993


        We have had no problem re-using Northern (or Southern) blots on either
Hybond N+, Gene Screen Plus or Nytran Filters.  We strip them by boiling for
5 min or so  either in dH2O or 0.5% SDS.  We cross-link the blots with UV just
after blotting (200uW/cm^2 for 13 min).  We have been able to re-use our blots
4-5 times without any problem (signal strength does fall a little @ each use).
Perhaps your problem lies in cross-linking the RNA to the filter and when you
strip the blot everything goes.  We have never used the alkaline fixation
method for Northern/Southerns so I can't tell you if that is inferior to the
UV cross-linking or not.
        Hope this helps.
================================================================================
Melissa Melan                                   mmelan at lucy.wellesley.edu
Department of Biological Sciences
Wellesley College
Wellesley, MA  01281
================================================================================



More information about the Methods mailing list