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    • #9623
      samkh918
      Participant

      Hello,

      I have used 30ml KIMBLE test tubes for DNA MaxiPrep application. I still have traces of DNA on the sides of the tube and I have tried washing it with high pressure water or leaving water in it overnight for the DNA to redisolve but none have worked and I still see the traces on the sides.
      Should I use a brush or is there other more efficient means? Is there any better solution than water to redisolve the DNA?

      Thanks for your help

    • #84036
      MrMistery
      Participant

      trizol?

    • #84054
      canalon
      Participant

      Tris 10mM at pH7.5 might be a little better, DNA do not resuspend too well in acidic solution.

      But what do you want resuspend the DNA or simply degrade it. In the latter case a 0,1M HCl solution should do the trick. It might not resuspend the DNA, but it will be depurinated and will not interfere with any other experiments. And brushing or using a dishwasher could also be really efficient (in fact some lab dishwasher have an Acid rinse in their cycle).

    • #84063
      snowcapk
      Participant

      Since it’s a maxiprep, I don’t think the OP wants to degrade the DNA, but rather to collect it.

      I have had this problem before. If it’s a large construct like a BAC that you are purifying, *do not vortex* to resuspend. Try heating the solution at ~37C for 20 min. This should cause the DNA to resuspend in solution. You may still see a white pellet there, but it’s likely that the majority of the DNA has resuspended at that point, so you might be able to give up on what remains.

      Try measuring the concentration of DNA in the supernatant as it is. You may find that you have plenty of recovered DNA in solution.

    • #84064
      samkh918
      Participant

      Thank you for your feedbacks.

      Yes, it’s after the MaxiPrep, so I have already collected my DNA and just want to get rid of these residues on the sides of the tube so that they won’t contaminate my future experiments.
      I will try the brush and detergent and if that’s not successful, I will use the 0.1 HCl.

      Thanks again for your help.

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