DNA GEL ELECTROPHORESIS

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    • #11071
      amberdagp
      Participant

      Hi can you please help me…
      The question i have been asked in after doing my Electrophoresis practical is: measure how far each of the observable fragments move (mm), tabulate the data and constrct a fully labelled calibration curve of migration distance against log10 (fragment size)

      Firstly, how do the measure the fragments, can anyone please tell me this very clearly as i do not get it, what parts do i need to measure and what i need to show in my table…

      Thank you very much in advance
      take care

    • #89726
      canalon
      Participant

      You need to measure the distance between the wells and the observed bands. Tabulate
      band -size -log10 fragment size -migration in mm
      use the table to draw the curve.

    • #89742
      amberdagp
      Participant

      Ok thank you 🙂 by the way, to measure the lanes where the fragments are undistingushable, and all you can see is a big shaded area in the lane, how would you measure in mm the distance of migration? this is as i need to get the average distance of a lane where the fragments are not separate and are all bunched together

    • #89743
      amberdagp
      Participant

      also why are base pair fragments different in number with haeIII in comparision to hindIII??

    • #89749
      canalon
      Participant

      If your fragments are not separated, I can’t do much. But you should measure to the front of the band. And different restiction enzyme have different cutting site, so likely different fragment size: think of divinding a book in fragments everytime a certain word appears, say "car" for one and "sleep" for the other, you would not expect the different fragments to be the same size. It is the same with restriction sites.

    • #89761
      MrMistery
      Participant

      ^ nice analogy patrick

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