dissolving DNA - (Mar/19/2007 )

hi evryone..
is it okay if i dissolve my DNA pellet in TrisCl( pH 8) without EDTA..?will it affct my subsequent reactions anyway?
what is the best method to dissolve DNA pellet after plasmid prep.?

that is okay.

However without EDTA, long term storage of your plasmid is jeopardised. There is a vastly increased chance that your plamsid will degrade from contaminating nucleases (already present of accidentally introduced at some later time). But if the DNA is for immediate use, and you care not for long term storage then it shouldn't matter.

As for down stream reactions, enzymatic manipulation (especially ligation) is improved without EDTA. (EDTA is inhibitory, unless diluted out).

Best way of dissolving DNA pellets?
Well, it would be to make sure that the DNA was very clean, then add TE (for my midiprep DNA). Sit back and watch an amazingly large pellet of DNA skips across the water's surface before vanishing into solution. That is the best way of dissolving DNA pellets... zero effort and a smile of satisfaction of job well done.

else

incubate DNA for 10min at 68 Celsius. If the DNA is moderately clean, it should go into solution.

I agree with perneseblue. EDTA works as metal chelator... meaning it will actually stabilise any DNA. (I am not too sure.. I think it is correct, can someone clarify it?)

I know few solutions to dissolve DNA pellets. One is the TE buffer (always get the one with reduced EDTA as EDTA might be potential inhibitor for subsequent reactions). Second is the NaOH and neutralised with HEPES (This will be easiest to solubilise the pellet). Final one is just water.