Cloning cDNA directly from PCR products
gruther at bilbo.bio.purdue.edu
Tue Mar 10 14:44:19 EST 1998
In article <01bd4873$ba1f6320$5d227681 at chm407-1.chem.ttu.edu>, "Benan
Dincturk" <knben at ttacs.ttu.edu> wrote:
> Christophe Jacob <jacob at scbiol.u-nancy.fr> wrote in article
> <jacob-0503982016450001 at morchella.scbiol.u-nancy.fr>...
> > Hello,
> > I'd like to clone several PCR fragments into plasmid vector. Their size
> > ranging from 200 to 700 pb.
> > Is it possible to use them for ligation after a step of purification (
> > PCIH, CI) and precipitation then digestion with the restriction enzyme,
> >* without electrophorezing them on agarose gel.*
Nipped and tucked and edited for emphasis
If you don't separate the digestion bits from that which you wish to
clone, you're going to have background probs unless you have a probe that
will allow you to screen by colony hyb.I'd take the extra couple of hours
to perhaps save myself several extra days.
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