You're correct: genomic DNA does get flash-denatured in a miniprep. That's a feature, not a bug! :)
A miniprep is intended to collect small, supercoiled DNA rings, which are able to resist this denaturation because of their supercoiling, so they can be collected in the supernatant fraction at one stage of the miniprep, leaving most genomic DNA behind.
If I recall correctly, the denatured genomic DNA is also effective at helping to capture some of the protein fraction, so it has the side effect of helping reduce that also.
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Dec 4, 2020, 12:18 by andreas.t.stuermer@gmail.com:
Hi everyone!I was wondering the following:During a miniprep, you first lyse the cells with sdoium hydroxide and then neutralize everything with aetic acid.Now a question came up that I never thought about: Will this not denature the DNA? During the high pH of NaOH there will be a lot of OH- groups floating around interacting with hydrogen bonds?Will the DNA not be partly or fully single-stranded then?Will it fully reaneal very fast, or not? will there be non-covalently joined and partly overlapping plasmid strands?Not that it really matters for routine applications but I find it very interesting and would be interested if anyone has thought about this, knows literature about it or knows the right keywords.Thanks!
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