IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

Charged vs. uncharged nylon membranes?

Stephen Perry bafq5 at central.susx.ac.uk
Fri Nov 10 07:06:17 EST 1995

ssilverstone at csubak.edu wrote:
: A question:
: I'm dot-blotting total bacterial nucleic acids then probing with DNA
: probes complementary to the 16S rRNA. The protocol I'm looking at
: specifies uncharged nylon membranes, but the Amersham literature says that
: their Hybond N+ is the best membrane for RNA blotting (positively charged
: nylon). Does anybody know why charged or uncharged nylon might be
: preferred for probing RNA dot-blots with DNA probes?  I am planning to
: label the probes using the Amersham ECL kit.-Sara


I have used N+ in the past, but always found there was a backgound
problem. For dot blots I would be particularly sceptical as you have
no separation of RNA species so you cannot judge whether the probe is
hybridizing specifically. I only used it for a couple of Northerns and
gave up when just about everything + the markers lit up!! I did not take
the time to alter the hybridization conditions or probe conc or washing,
so you may have more luck.

good luck  Steve Perry

More information about the Methods mailing list

Send comments to us at biosci-help [At] net.bio.net