doubt regarding agrobacterium DNA restriction digestion

Michael J. Prigge via methods%40net.bio.net (by mprigge from indiana.edu)
Wed Jun 18 14:29:50 EST 2008


What are the symptoms?  Are you trying to digest plasmid DNA?  If you  
can see the DNA on a gel but it looks uncut, it might be due to the  
differences methylation patterns.  I don't know what sequences are  
methylated in Agro other than GANTC, but you should be able to find  
that info somewhere then check to see if your enzyme might be affected  
by that methylation.  If you are not seeing anything on the gel, it is  
likely because binary vectors are (with a few excpetions) present at  
low copy numbers.  I typically digest the plasmid yield from ~2 ml of  
culture for each lane on a gel.
Mike
> hi all
> as i am attempting to digest agrobacterium DNA i am not able to do  
> that.
> please help me regarding this.
>
> -- 
> prasanthi


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