In-Vitro mutagenesis using DNAse I

William Guy Fairbrother wgf3 at
Thu Aug 6 11:41:54 EST 1998

I wanted to generate a series of small deletions in a large RNA
element to probe which parts were necessary for function. I was thinking
of using DNAse I to linearize plasmid  and by polishing the ends and
religation, create a series of small deletions? has anybody ever heard of
I was thinking of using Mn++ as the co-factor but I heard that only gives
1-2 bp overhangs. What would Mg++ give (somebody said approx 12bp
overhangs) . As you can see , I'm in the dark. if anybody has done this or
heard of it. Please let me know. thanks a lot

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