Hello
Does eukaryote DNA extraction need to perform in sterile condition with sterile (TE) buffer?
And what about plasmid extraction?
I read somewhere that plasmid DNA extraction does not need to be sterile as plasmid DNA is supercoiled and remains in solution when sodium acetate is added. This causes proteins and chromosomal DNA to precipitate and allows for the extraction of the purified plasmid DNA.
Solmaz Morovati not need for sterile condition for plasmid extraction
Depending on the intended use of the DNA, it might be necessary to filter the DNA with a sterile filter (0.2µm) e.g. before you use it for transfection of eukaryotic cells. Other than that there is no need for sterile buffers because plasmid DNA is mostly stored in the freezer. E.g. if you use it to transform bacteria I guess using sterile DNA would make little sense.
You'll want to use sterile/certified DNA/RNA free tubes and pipette tips. But in general, you often do not need to work in a sterile environment. Although, depending on the type of work you do in your lab, there could be risk of DNA contamination from other sources.
Hüseyin Besir Thank you but filtering DNA reduces the amount of our sample. So, isn't better to do the extraction with sterile solutions?
As April Armes mentioned microtips and eppendorf tubes need to be sterilised but not necessary the working area.
Solmaz Morovati Using sterile solutions only does not make the final product completely sterile as long as you can't perform the final step under sterile conditions. As I said, it depends on how you want to use the DNA. For use in cell culture I would recommend sterile filtering or a final step under sterilizing conditions.
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