Digestion near the end of the DNA

BENNY SHOMER 9238 pc386 at ccsg.tau.ac.il
Thu Aug 19 10:25:11 EST 1993


Hi All.

Vivian Miao (vmiao at oregon.uoregon.edu) wrote:
: Has anyone had experience cutting EaeI or XmaIII (EagI) sites close to an
: end of DNA?  Anyone had GOOD cutting of NOTI when there is LESS than 7
: bases between the site and the end of the DNA?

: I know about the useful references in the backs of various catalogs (e.g.
: NEB,  Stratagene), and of one paper in Biotechniques, but I can't find any
: information of these enzymes specifically. [Del, Del, Del....]...

We had some good experience with doing this indirectly.
The fragments to be cut are ligated into concatamers (Fill-in if needed...).
The concatamers are then digested with your R.E. of choise, which now cuts
within the molecule with no problems. You should have enough fragments cut
correctly.

Good luck, Benny.

Benny Shomer.
Tel-Aviv University, Israel.
pc386 at ccsg.tau.ac.il



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