methods to prevent digestion by REs?

Basavaraju Shankarappa bsh at MED.PITT.EDU
Fri Apr 22 09:07:43 EST 1994


Hi folks:
	We have a situation with one of our clones where we thought the 
restriction site was an unique one, but now it turns out that there is an 
extra one in the vector.  Now we need  a method that will prevent 
digestion by restriction enzyme at one site while allowing it to cut at
the other site.  
	I remember seeing an article or a note somewhere 
which said that it is possible to use an oligo as a band-aid on the sequence 
where you want to avoid digestions by the RE.  More like forming a triple
helix at the position.  Does anyone know of the full reference of this article
or opinions concerning how efficiently does the digestion get blocked.
If you have any new ways, that this can be done, I would appreciate that too.
I also thought that one can use an overlapping methylase recognition sequence 
to block some of the sites.  Please correct me or guide me.
I can use any help.
TIA
Raj Shankarappa
bsh at med.pitt.edu  



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