Nylon membrane blues

Julie Wilberding jwilber at pop.nd.edu
Thu Nov 17 11:59:49 EST 1994


I am having problems getting good sgnals on DNAse I hypersensitive assays. 
I suspect that it may be blotting.  I alkaline blot onto Nylon membrane
from biorad (zeta probe).  Is there anything about the gel (1.4%), or DNA
sample that might be causing this.  Even my parent fragment is coming up
light.  I load 20 ug of DNA per lane.  After blotting I neutralize for 10
minutes in 50mM Tris pH 7.5 and then bake for about 45 minutes.  I AM
REALLY DESPERATE!!!!!

Can anyone offer any clues or advice?  Thanks!

Julie



More information about the Methods mailing list