Partial RE digestion - please advise!

Victor Levenson U10784 at UICVM
Sun Feb 11 17:57:08 EST 1996


Hi, bionetters;

I have a problem with a partial digestion and I need your input.

Plasmid has 2 sites for BsmBI, one of them in the fragment that I need.
I tried to get a partial by modifying:

1. plasmid:enzyme ratio
2. temperature
3. time of the digestion


I can get a partial, but the majority of the cuts are at the wrong site; I 
get a lot of uncut stuff, a considerable amount of linear form and some 
double-cut fragments, second digestion (with another enzyme) reveals that most 
cuts are at the wrong site.  Today I tried digestion in the presence of EtBr - 
works great for DNaseI - and I could not get ANY cuts!

Is there any way around this crap? 

Any hints will be APPRECIATED

Is there a 



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