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