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partial digestion - match partial digestion conditions (Dec/12/2004 )

Hello!
Has anyone ever performed partial digestion of DNA, I try to cut DNA this way, but it is hard to match the conditions. Maybe there is some clever method to make it efficiently.
Regards
Tomek

-tomgar-

There are some old tricks.
If you add ethidium bromide to the restriction digest, the enzyme will make a single cut and stop. I don't remember the exact concentration of EtBr required.
Another option dates back to the old days of cesium gradient purification of plasmid DNA. The preferential initial cut by a restriction enzyme can vary depending upon whether or not the DNA has been run over a cesium gradient and then phenol:chloroform extracted. The same might be true of DNA that has been prepared by alkaline lysis and then plus or minus purification by one of the numerous commercial kits.

-tfitzwater-