We have isolated DNA with the Qiagen Powerlyzer Powersoil Kit (with addition of Proteinase K) for NGS downstream applications (16SrRNA/ITS2) and encountered the problem that the isolated DNA is not stable at -20°. After storage of some weeks and thawing, gel electrophoresis after PCR amplification barely shows any bands or only smear. The longer the DNA was stored at -20°C the worse it gets until the complete loss of DNA.
According to the manufacturers protocol, the DNA is to be eluted into provided 2ml collection tubes, whom we assume they should be suitable for DNA storage, or?
We were thinking about the suitability of the C6 solution (elution buffer) for DNA storage or maybe the quality of the collection tubes, but we couldnt find any hint so far.
Any suggestions or has anyone also experienced this kind of problem?
It seems to be a problem of the Powerlyzer Powersoil kit AND also the fungal DNA itself. The bacterial DNA still gives better PCR results after storage and thawing than the fungal DNA. But why is that?
We appreciate any suggestions or/and solutions for this problem!
I have had a few similar drawbacks with the equipment but not enough to describe its DNA isolates as 'unstable'. It could be a function of the nature of the DNA molecule itself, that is, something in the reaction mixture or receptacle just does not agree with it. And that is what we are missing.
Respected Anna Lara Ernst , try eluting lesser volume. DNA concentration may be less.
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