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Salt precipitating out with my DNA

DNA extraction precipitation

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#1 RhiRhi

RhiRhi

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Posted 23 May 2013 - 09:07 PM

Hi There,

I hope someone can please help me, I have 800 buffy coat samples that need DNA extraction.

I have been using a salting out method for 7 years and have never experienced any salt issue.

Basically, when I spin down my sample at 4 degrees, not all the salt is spinning out. So when I tip off the supernatent and add 100% ethanol, the DNA precipitates out, but so does the salt. This then becomes a rescue mission to capture the DNA before it sinks to the bottom and is enveloped into a pile of salt. I have managed to rescue them, but they are not happy DNA precipitates. They are white, and hard/dense, not viscous like normal DNA. After plenty of stress and washing them in 80% Ethanol 5 times, the majority turned out OK.

Problem is I can't go through this for 800 samples...

Seems like it is an isotonic/salt imbalance problem. I have tried many different ratios and volumes and just can't fix this problem. AAAHHH!

Please HELP!

Thanks!





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