oligo cloning

ChenHA hzhen at freeuk.com
Mon Feb 7 16:16:21 EST 2000

"R. John Lye" wrote:

> Caroline Szymeczek-Seay wrote:
> > Vector is cut with SmaI and CIPed.  Oligos are kinased.
> Be careful of over-CIAPing - in my experience, too much enzyme and/or
> too
> long a reaction will damage the ends of the DNA.  Make sure that all the
> CIAP has been removed, also.  I usually use a little SDS/proteinase-K
> digestion followed by phenol/chloroform to be sure it is all gone.

One useful protocol for blunt-end ligation without CIP:

but replace EcoRV with SmaI, pBluescript with pGL3 in this particular case.

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