Cutting the ends of dna

Rafael Maldonado rmaldonado at
Tue Nov 2 09:48:03 EST 1999

Aidan Weatherill wrote:
> If someone can suggest ways of increasing the efficiency of a) cutting at
> the ends of molecules and b) doing triple ligations, I would be very (very)
> happy

Cutting at the edge of the DNA molecule is tricky, and it depends on the
enzyme you are using. The New England Biolabs catalog gives some good
information on this in one of the appendices (not relation with the
company). SacI it is quite bad for this, and BamHI is not very good. I
would suggest adding extra bases (from 3 to 5-10) to the end of the
oligo for improving the cutting. Check the NEB list for choosing better
enzymes for this job.


Rafael Maldonado
Divison of Genetics
University of Alicante (Spain)

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