Problems with restriction digests

Michael Rosemann rosemann at gsf.de
Tue Jan 27 10:41:18 EST 1998


>We are having problems with our restriction digests that I am hoping someone 
>will be able to solve.

I had a similar problem with a plasmid that was kept in TE buffer. If to much of the
TE buffer is transfered to the RE reaction will cause the EDTA to bind most of the 
Mg++ ions, which the Restriction Endonuclease needs for normal activity.
I could get rid of the problems by precipitating and redissolving the DNA in pure water.
A colleague, however, recommended to simply add some extra MgCl2 to the 
RE buffer to compensate for the EDTA effect.

Greetings  M.Rosemann (rosemann at gsf.de)



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