Protocol for plasmid-miniprep

Susanne Rohrer "srohrer(removethis)" at immv.unizh.ch
Fri Aug 4 06:46:17 EST 2000



pairan at student.uni-kassel.de wrote:

> Hello everyone!
>
> I`m searching for a 'fast' plasmid-miniprep-protocol for
> Enterococcus sp.
>
> Is there anyone out there, who has a script to prep them in
> ca. 5 h?

even less, if you have an enzyme that can lyse the cell wall. for S.
aureus, we add lysostaphin to the resuspension buffer of a commercial
miniprep kit, (macherey-nagel, quiagen etc, no affiliation) and incubate
at 37°C until it becomes clear (about 10 min). then we lyse and proceed
as for e. coli.

depends on how large your plasmid is, they all don't elute well if it's
over 15 kb.






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