Why is my second DNA elution have lower 260/280 ratio?

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Why is my second DNA elution have lower 260/280 ratio?

Postby chunchunmaru » Apr 16 2017 9:25 am

Hi,

Can anyone explain this: how/why does a second elution of DNA have a lower 260/280 ratio (and 260/230 too!)? Also, why is the first elution really bad; smears are very prevalent. The 2nd elution always has nice and high MW bands with very little smears. But, the problem is that the 2nd elution has low ratios: <1.5 for both ratios. And what puzzles me more is that 1st elu looks horrible in gel but really pretty in spectro (~1.8 and 2.0 - 2.2), plus high DNA quantity.

I am using a column-based, commercial extraction kit and I will be using the DNA for SNP genotyping using Sequenom. Nanodrop for spectro measurement. Any ideas?

Thanks!
chunchunmaru
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