Partial RE digestion - please advise!
U10784 at UICVM
Sun Feb 11 17:57:08 EST 1996
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
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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