The space between two RE sites?

bhjelle at carina.unm.edu bhjelle at carina.unm.edu
Mon May 10 09:53:59 EST 1993


In article <1sjtilINNq77 at umbc7.umbc.edu> xlin at umbc.edu (Xiaoying Lin) writes:
>Hi, all
>	I am planing to make an linker with 2 RE sites in it. The double cuts 
>will be needed for subcloning. Any idea on how far away they should stay away 
>from each other? 	Thanks.
>
Check the New England Biolabs catalog (great cover, NEB!) for
a table in the back concerning the efficiency of various RE's
at cutting near the terminus of a DNA sequence. RE's vary
a lot in this respect. Of those I am familiar with (School
of Hard Knocks), Hind III is not very good at cutting near
the terminus but Kpn I and Xba I are. If you have one enzyme
that is good at this and one that is not, just make sure to
make the first cut with the bad actor, and the second
cut with the good enzyme. I would not, in any  case, space
the sites less than 3-4 bases apart.

Good luck.

Brian



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