dna purity is 17? what does it mean?

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    • #15995
      genetherapy
      Participant

      Hi everyone,
      I’m constructing a new vector and before starting ligation I measured dna purity of my gene. The dna purity that I measured is 17. More more than 1.8 😕 What does this mean, is my dna is so pure from proteins etc 😀 I measured this three times, it is unable to understand for me. Could anyone explain me this?
      Thanks for yor help

    • #109236
      JackBean
      Participant

      are you sure it’s 17 and not 1.7? How did you calculate that?

    • #109243
      genetherapy
      Participant

      Yes I’m sure that it is 17 not 1.7. The spectrophotometer calculates automaticly when I enter my dilution ratio. I used spectro lots of times and that is the first time ı see a value like this. But ı make dna cleaning and concentrating prosedure and forget my dna sample 5 or 7 minutes in the final step(elution step). Is this the matter I really don’t undestand??

    • #109244
      genetherapy
      Participant

      I mean ı forgot my sample to centrifuge for about 5 or 7 minutes besides only 1 minute waiting.

    • #109246
      JackBean
      Participant

      you’re centrifuging it for 5 minutes?

      Anyway, what was your concentration? Such out-of-mind values usually come up when you have very low concentration and thus very low absorbance.

      BTW with the 260/280 ratio higher than 2, the DNA is contaminated with some RNA, phenols or something like that. But I doubt anyone ever saw 17 😆

    • #109252
      genetherapy
      Participant

      Interestingly my dna concentration is 157,07 mikrogram/ml 😯

    • #109254
      genetherapy
      Participant

      I also want to ask another question like this problem. Today I purified DNA from pellets. Then I read their purity and concentrations in spectrophotometer. The purities range between 1,8-1,9. Really good and concentrations are good also. Then I digest this samples with an enzyme and look if it digest, the digestion was perfect. I cut the bands that I want and the problem starts at this point: I made gel extraction and now the purities range between 3,5-5 . The concentrations are 5 mikrogram/ml. I’m so unhappy that you can’t guess after the perfect purity and concentration from first step. Is it the agarose that wastes my samples? What will I do?

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