restriction map – gel

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    • #9310
      spinkster
      Participant

      Hi guys,

      I have done a restriction map gel and got what I expected, but my uncut pUCPH seemed to seperate into multiple bands.
      (I was only expecting 1 band as surley no restriction sites used therefore surely it should all be one size)

      does anybody know why this is?

      any help would be much apprieciated

      Matt

    • #82909
      Cat
      Participant

      To get a single band, you need to cut your vector with a single cut enzyme. Uncut plasmids can be a mixture of topologically different forms (nicked, linear, covalently bonded, single stranded circular, and supercoiled) and none of them will migrate as expected of the cut linear vector.

    • #82923
      spinkster
      Participant

      Cheers for your help cat 🙂

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