I need to clone a 34kb insert into E.coli. Suggest good vectors. IS PGEM 3z good...
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Posted 06 March 2011 - 03:17 AM
will the 34 kb fragment contain prokaryotic DNA that is possibly active in E. coli?
I would certainly go for a low copy vector that is propagated stable. I would suggest vectors with an F-plasmid backbone or mini-F backbone (like one of the first pBAC108L). The pBeloBAC11 is available from NEB ...also a good choice since it allows for blue/white screening. You can also try looking for PACs ...but i do not know where to obtain them.
A good overview on that topic can be found on openwetware.
Regards,
p
If the fragment is of other origin than prokaryotic go for a Bacmid
I would certainly go for a low copy vector that is propagated stable. I would suggest vectors with an F-plasmid backbone or mini-F backbone (like one of the first pBAC108L). The pBeloBAC11 is available from NEB ...also a good choice since it allows for blue/white screening. You can also try looking for PACs ...but i do not know where to obtain them.
A good overview on that topic can be found on openwetware.
Regards,
p
If the fragment is of other origin than prokaryotic go for a Bacmid
krithi, on 06 March 2011 - 01:42 AM, said:
I need to clone a 34kb insert into E.coli. Suggest good vectors. IS PGEM 3z good...
