BamHI Without Star Activity

Steve Rodems smrodems at students.wisc.edu
Wed May 18 23:11:30 EST 1994


In article <1994May19.005402.28015 at vaxkiller.agi.org>,
vaxkiller.agi.org!rlynch (rick lynch) wrote:

> I am having some trouble getting a clean BamHI digest of a plasmid without any  
> extra bands.  I am using 5X excess of enzyme by units, and am using a standard  
> buffer with 150 mM NaCl.  It is important that I get only linear plasmid, and 
> I can't gel purify the fragment for other reasons I won't get into now.

My first suggestion would be to use BamHI from another company than the one
your using.  I have never had problems with BamHI from BRL.  This enzyme
cuts in REact 3 which is 100mM NaCl.  Maybe you buffer is too high in salt.
 I have never used BamHI from NEB but several other enzymes from NEB work
well in my hands so they may be a good source as well.

-- 
Steve "Some day I will get the hell out of Wisconsin" Rodems

"Then I am here for the Lee family renioun ... shur-wajo-shur"



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