cloning of small DNA fragments (66 bp)

Frank O. Fackelmayer Frank.Fackelmayer at uni-konstanz.de
Thu Aug 5 10:49:32 EST 1999



Frederik Boernke wrote:

> Gregor wrote:
> >
> > ...or just forget about the template - it´s much larger than the insert
> > and will be cloned only in a few cases. this you can check with colony
> > PCR.
> >
> > Gregor
>
> I only thought because of the selection marker which would be the
> same in donor and target plasmid. And even though the template
> is very dilute it will make out the plethora of recombinants.
>
> Ricky

Hi Ricky,
You are absolutely right to consider the template a problem in cloning PCR
products. I reproducibly got a much higher percentage of right clones after
DpnI treatment. Therefore, I do it routinely now (or gel-purify the fragment
before ligation).

Frank





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