I want to precipitate pBR322 after treating with peroxynitrite so that i can do restriction digestion further.If anybody have any protocol for this please share.
Done with the same method but I am not getting it....After treating with peroxynitrite I was adding 3 volume of chilled ethanol n kept it for overnight incubation at -20 degree C. Next day centrifuging at 120000 rpm for 20 mins and again wash with 70% ethanol.But stll i am not able to get.
I want to generate single stranded nick in pBR322 by peroxynitrite. Nick generated by peroxynitrite is confirmed by running the DNA on agarose gel.So i want to precipitate DNA after treating with peroxynitrite.
A normal PCR purification method or Ethanol precipitation should work. Are you using kits ? if yes, you can try a kit used in plasmid purification or PCR purification, that might work 'cause any silica based column would trap your DNA and separate it.
And, is there any specific reason you want to induce ss nicks in the plasmid ?