Phenol/CHCl3 rendering dsDNA single-stranded

Gerd gerdn at ibg.uit.no
Wed Dec 2 07:22:18 EST 1998


What about dephos, (and maybe phen/chl. extr.) before the gel
purification? The the strange stuff wouldn't be present in your
ligation.

Gerd

R.N. Leach wrote:
> 

> I cut my plasmid, gel purify it by cutting the appropriate band out of an
> agarose gel and eluting the DNA by centrifugation. After EtOH/NaoAc
> precipitation and resuspension I dephosphorylate and then phenol/chloroform
> extract the cut, dephosphorylated plasmid DNA ready to ligate in my insert.



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