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Ammonium acetate DNA extraction mistake - (Oct/27/2011 )

Hi all,

I am using an ammonium acetate extraction protocol to extract DNA from some insect samples. However, instead of adding Digsol to the chopped up sample, I made a mistake and added ammonium acetate (this should have been done in the step after digestion!). I panicked and added the digsol anyway, so now I have a tube containing the sample, digsol, ammonioum acetate and proteinase K currently incubating at 55oC.

Does anyone know whether I've messed up my extraction by doing this and if so is there a way of rescuing it? Or can I carry on with the protocol as normal (i.e. leave to digest overnight, then add more ammonium acetate, then centrifuge and carry out the ethanol washing steps)?

Any advice would be appreciated!



No idea if this works or not, but Proteinase K is quite stable and should survive this. At least I'd check the pH and adjust it to the pH of the extraction buffer to avoid a too low value.