plasmid wanted

Alex Brands abbrands at artsci.wustl.edu
Fri Nov 2 17:00:14 EST 2001


On 2 Nov 2001, Johannes Graumann wrote:

> Cornelius Krasel <krasel at wpxx02.toxi.uni-wuerzburg.de> wrote:
> > You will be disappointed, but I am using good old Bluescript cut with
> > EcoRV for this case.
>
> I was also browsing around for "standard" vectors that would do this
> but as is the case for
> Bluescript, they have all the common re sites around in the mcs and
> wasn't allow easy evaluation
> of PCR introduced sites.

Why can't you just cut out the mcs on either side of EcoRV?

> Yep, got it right: I need a vector to subclone PCR products with primer
> introduced restriction sites. Don't want to do TAing - strongly believe
> in PfuTurbo.

You can treat PfuTurbo products with non-proofreading Taq (no cycling,
just incubate at 72C) to add the overhang for TA cloning.

Alex






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