in situ cracking?

Neil Saunders s3003334 at pop3.unsw.edu.au
Tue Jul 25 00:09:21 EST 2000


> I'm sure you'll attract the normal protocols for cracking/plasmid
> sizing so I will not reiterate them here HOWEVER an ex-colleague
> of mine claims he has heard of a protocol in which you cast the
> agarose gel with some kind of detergent in it.  You can then add
> bacterial suspension directly to the wells for "cracking in situ".
> I've searched various protocol lists but have never found this one.
> Has anyone else heard of it as it sounds very useful?

I once used a similar protocol to this for analysis of megaplasmids in
Paracoccus denitrificans; basically cells were added to the well plus
detergent (I forget which one, something non ionic), incubated for a while,
then sealed with agarose.  The idea was to avoid shearing the megaplasmids.
I'm not sure why anyone would want to analyse 'normal' cloning vectors this
way, but I could probably dig out the protocol if anyone wanted it.

Neil
--
School of Microbiology & Immunology,
University of New South Wales,
Sydney 2052,
Australia

Ph: +61 2 9385 2093
Fx: +61 2 9385 1591
email: neil.saunders at unsw.edu.au
http://www.crosswinds.net/~nfws/index.htm
ArchaeaWeb
http://www.crosswinds.net/~nfws/archaea/index.htm








More information about the Methods mailing list