enzymatic cleavage of DNA to small pieces (20-40bp)?

Dr. Duncan Clark duncan at genesys.demon.co.uk
Wed Apr 29 04:35:20 EST 1998


In article <3545BC75.D2256D0 at chclu.chemie.uni-konstanz.de>, Frank O.
Fackelmayer <fof1 at chclu.chemie.uni-konstanz.de> writes
>I´m currently thinking about enzymatic cleavage of DNA to small fragments,
>preferentially between 20 and 50bp. Ideally, cleavage should create blunt ends
>that work for cloning.  So, DNase I or micrococcal nuclease are not the
>enzymes of choice, also because they don´t stop digestion until the DNA is
>broken down to TOO short fragments (1-4nucleotides each).
>What I would like to use is a kind of restriction enzyme with a 2bp (or 3bp)
>recognition site. I remember that I saw a paper some time back where the
>authors used something like that, but I don´t remember the details (not even
>if I saw the paper published or it was one I had for review...)


What you want is CviJ I but it is very very expensive. It recognises
RG/CY and has a star activity so it cuts with even less specificity
under the appropriate conditions.

Try looking at REBASE on NEB's web site for who sells it.

Duncan 
-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
TEl/FAX 01252376288
http://www.dnamp.com
http://www.genesys.demon.co.uk



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