Use of positively charged nylon membranes for library screening
daram at ENSAM.INRA.FR
Thu May 29 08:03:33 EST 1997
I am in the process of optimizing probe/hybridization conditions on
Southerns for subsequent phage library screening and I have just
discovered that the membrane I am using (ICN BIOTRANS+) is not
recommended for phage lifts as it is positively charged.
Has anybody used either this or other positively charged membranes in
library screenings? If so was any change in methodology required, i.e.
from traditional denat, neut, 2xSSC, for carrying out the lifts.
One person I have spoken to suggested that there might be a problem with
increased background due to proteins being bound by the +ve charge, as I
am hoping to be able to reprobe the membranes 2-3x can anybody verify
whether this is likely to be true.
thanks in advance for any help/suggestions
Logan at ensam.inra.fr
More information about the Methods