I have extracted DNA from a vaccine containing aluminium hydroxide gel using sodium citrate (tribasic). The extraction has work well and I have a good qubit reading for dsDNA as well as a good nanodrop reading of 260/230 and 260/280. However, when I run a PCR on the sample, the internal positive control does not amplify indicating inhibiting factors (I suspect aluminium ions). I have tried diluting the sample but still no luck. Has anyone used a good protocol for removal of inhibiting metal ions?
Many thanks in advance
Hello Zayed,
you might consider agarose gel purification. After running the sample you can excise the DNA and use a gel purification kit to remove the agarose. Otherwise silica-based columns used for DNA preparations with one or two additional washing steps might also do the trick.
Cheers, Christian
Aoa dear, you can remove A ions by adding Na or K nitrate dil. solution to precipitate out Al ions.
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