I have used to Qiagen powersoil pro kit to extract some DNA from bacteria, my first bunch of samples had good concentrations between 200-300 ng/ul on Qubit, and 300-400 ng/ul on nanodrop for comparison. The 260/280 ratio was 1.8 or above for all but 260/230 was 1.2 or lower.
I read about adding in an additional ethanol wash during the extraction so I did this on my second bunch of samples. 260:280 values were 1.8 or above, 260:230 values 1.8 or above, so this was good. On the nanodrop I got concentrations of 500 - 600 ng/ul, however the Qubit couldn't read these samples and said it was 'TOO HIGH'.
Can anyone help me understand what's going on with these samples? I have about 60ul of sample left so am trying to find a clean up protocol for them (maybe isopropanol?), but the second bunch of samples I do not know what to do?
Thanks for your help.
Low 260:230 would indicate chemical contamination. You can try to precipitate all your extracted DNA with ethanol, or for new extractions you may want to use higher elution volume and water instead of elution buffer
From all of your extracted DNA make precipitation with ethanol, or for new extractions you may want to use higher elution volume and water instead of elution buffer
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