I am using the 30-uL QIA gel purification kit, and each time I test the product on a nanodrop after, the 260/230 is in the 0.01-0.30 range. Subsequent reactions, like ligations have issues with extremely low efficiency. I typically elute with 30uL H2O. Asking around has lead me to believe that this means there is too much salt in my samples.
A Zymo Clean&Concentrate protocol afterwards typically works, but there's inevitable some sample lost when adding in extra steps like this.
I've tried all the usual recommendations (extra binding buffer wash, let wash buffer rest a few minutes at RT, extra ethanol wash, elute with warmed H2O), but nothing has worked so far.
Can anyone help me out with this?
Interesting question with great insights. Looking forward to the discussion!
I've honestly never had very good luck with Qiagen's gel purification kits, they always give me bad 260/230 ratios. I have no idea why. I've had better luck with NEB's gel extraction kits and I've heard good things about Zymo's also.
you have tried everything that I would have suggested so try a different isolation kit. Zymo since that seems to work for you. The excess salt may be keeping the sample on the column so a second water elution step might give a smaller amount but possibly salt free if yield is not the main factor
Interesting question with great insights. Looking forward to the discussion!
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