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Topic: DNA Depurination and Depyrimidination  (Read 7365 times)

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Offline ramma

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DNA Depurination and Depyrimidination
« on: August 26, 2011, 04:46:34 PM »
Hi,
I'm not sure this is the right board for this question, so sorry if it isn't.

I'm trying to digest DNA down to just the bases, and I'm having difficulties finding a protocol on how to. I was told by another student that a 88% formic acid boil at 175 C for 1.5 hours would remove both the purines and pyrimidines, and degrade the sugar, leaving the sample ready for TLC analysis. Yet this method is giving inconsistent results.

My DNA sample is just a direct extraction, but I do have DNA degradase plus that could be used before digesting to the bases.

Thanks for any help and I'll be glad to answer any questions.

Offline rjb

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Re: DNA Depurination and Depyrimidination
« Reply #1 on: August 31, 2011, 10:07:40 AM »
Ramma,

Sorry for the late reply, but have your tried 88% formic acid + 2% w/w diphenylamine? A 30 degree 18hr digest should do the trick...

Kind Regards

R

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