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LucaAster

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  1. I am using a UV spectrophotometer with DNA. If there is degradation of base pairs, would I expect to see a higher or lower absorbance value? There are more DNA strands after base pair loss, due to the strand being cut into pieces, but with fewer bases in total. So, the concentration will technically be higher? In that case, would the absorbance be higher also, or am I looking at this wrong and it only depends on the number of base pairs?
  2. Does anyone know of a simple procedure for isolating DNA from E. coli? I have considered using a freeze-thaw method with a freezer to lyse the cells, but I don't know if that would be most efficient. I have also seen some methods that where phenol is simply used to do it. If I did use the freeze-thaw method, would I need a buffer in the solution? How much bacteria should I put in the water to be freezed? For the DNA isolation, I wanted to use a centrifuge with equal parts of the cell mixture and phenol, but I'm not sure how long I would need to do that for. Is that the correct mixture to put in the centrifuge? After centrifugation, how would I separate the phase containing the DNA? Thank you!
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