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partial digestion


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#1 tomgar

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Posted 12 December 2004 - 11:56 PM

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

#2 tfitzwater

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Posted 15 December 2004 - 04:41 PM

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.




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