Xho I cutting problem

Jens Tornoe jt at neurosearch.dk
Sun Jan 16 14:44:48 EST 2000


Since "normal" restriction enzymes like XhoI cut palindromic sequences only,
it would not cut at CTCGCG. A too high glycerol concentration in your
reactions or use of the wrong enzyme buffer, however, can cause the enzymes
to cut at non-specific sites (star activity). Therefore you should never
have more than 10% enzyme stock in your reactions (NEB and other enzyme
manufacturers). Also, do not digest your DNA too long if you are having
problems.

Another explanation of your observation would simply be that you have
incorporated a XhoI site by an accidential PCR mutation. Sequencing your PCR
product can tell you if that is the case.

Jens

--
Jens Tornøe
NeuroSearch
Pederstrup, Denmark
http://www.neurosearch.com



<tunlu at med.wayne.edu> wrote in message news:3881F20E.3FD at med.wayne.edu...
> Hi:
> Xho I cuts CTCGAG.  Does it possibly cut CTCGCG?  We recently are using
> Xho I to cut our PCR products, but found that there was not desired
> band.  Any suggestions?
> yang
> ---






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