NEED PROTOCOL: PCRing genes from E.Coli

Jim Hill nospam at ima.org.sg
Tue Jul 4 03:48:31 EST 2000



Jim Hu wrote:

> Chris LaRosa wrote:
>
> > > Can someone give a working protocol
> > >
> > > for PCRing a gene from E.Coli genome?
> >
>
>
> Actually, getting PCR products from E. coli is pretty easy - for plasmids
> we just pick a colony into water in an Eppi tube, boil it, spin, take a
> SMALL amount of the sup and amplify.  If anything, using too much DNA can
> cause problems.  We've also done this for looking at lambda lysogens, so
> chromosomal loci should work too.
>
>

For plasmids I just dip a cocktail stick with the bugs on it into the PCR
buffer. Works every time.

Jim







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