No background plasmid

Hans Hartings isc2 at SPM.IT
Sat Nov 1 07:12:24 EST 1997


Dom Spinella wrote:

> > I've been looking for a plasmid that has a multiple cloning site in
> the
> > middle of a suicide gene, so that an insert is nessesary for the
> > transformants to survive. So far the only one that I've seen is one
> made
> > by Invitrogen, which I am told you can't plasmid prep yourself, but
> have to
> > by it from Invitrogen every time you want to use it.
> >
> > Has anyone heard of any other plasmids like this?
> >
> > Adam Griffith
>
> I assume you're referring to pZero plasmids from Invitrogen in which
> the
> MCS is in the middle of the lethal ccdB gene.  Maybe I'm missing
> something here but can't you purchase some plasmid DNA from them and
> then roll your own vector by just cloning any old fragment into the
> center of the MCS, preparing a large scale plamid prep from the
> recombinant, and then digesting away the insert as needed with the
> same
> enzyme or whatever flanking enzymes you'd like use for your real
> cloning
> work?

You don't need to do anything like that..... just grow a pZERO
containing host without IPTG and you can get as much plasmid DNA as you
like!

> Of course, there may be patent and licensing issues, but if you
> purchased the plasmid from them initially, who's to know that you made
>
> some more on your own?  Just a thought... -- Dom Spinella

   That's right: you can prepare the DNA but legally speaking you should
buy it!

H. Hartings





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