Plaque hybridisation with nylon membranes?

Steven Goldberg goldberg at bms.com
Wed Jun 5 08:19:25 EST 1996


In article <4p1erc$knn at sun4.bham.ac.uk>, c.hoege at bham.ac.uk (Carsten
Hoege) wrote:

> Hi there!
> 
> I'm up to screen a million phage plaques  of a genomic library in poppy. 
> I want to use square plates (25x25 cm). Since I haven't found a manufacturer
> providing nitrocellulose filters for these plates I thought about using
Hybond-N
> or Hybond-N+ membranes which come on a roll. Previously I used
nitrocellulose filters
>  with the standard screeningprocedure of Benton and Davis . Has anyone
experience, 
> how the different charge of nylon affects the screening, especially non
specific binding ?
> Thanks for any help!
> 
> Carsten

I have used Duralon membranes from Stratagene as well as Hybond-N+ for
phage library screening with excellent results.  Nylon is much easier to
handle, especially when using large filters.  The background is not much
worse than nitrocellulose...just follow the protocols that are included
with your membrane and do duplicate filters.

Good luck.

Steve



More information about the Methods mailing list