Most robust restriction endonucleases?

Harry J. Witchel Harry.Witchel at Bristol.Ac.Uk
Thu Apr 3 06:08:28 EST 1997


Felicitations Molecular Masters --
   I was wondering what your experiences are of the most robust
restriction endonucleases, i.e which enzymes seem to be least affected
by slight fudges in the buffer (eg TE in sample), impurities in the
sample, less than perfect pipetting, etc.
   For example: I find that Pst I will cut almost any time with little
or no partial cutting.  Pvu II is also very reliable for me under
unusual conditions.  I trust HindIII and Kpn I implicitly.  However, if
one of my samples is not perfect, EcoRI may not cut it, and if a less
than perfect sample is cut by BamHI, the result will have many
partials.  I cannot get BssHII to work under even the best of
circumstances.  Etc.
   What is your experience with the different REs?
Thanks,
Harry

Harry.Witchel at Bristol.Ac.Uk



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