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    • #10774
      lcronan
      Participant

      I am attempting to make a library of copy-number mutants. In order to do this, I wanted to use random mutagenesis to create the mutations. Does anyone know of a good, tried and true technique? I have tried a random mutagenesis kit, but so far have gotten no mutants. Of course, I may be doing something incorrectly, but I am not getting any helpful feedback from my boss… and yes, I have asked! If anyone has any advice, hints, knowledge, thoughts, etc. to share on the subject, I would be extremely grateful!

      Thank you!
      Leah

    • #88569
      mith
      Participant

      what kit?

    • #88593
      lcronan
      Participant

      Stratagene’s Random Mut. Kit

    • #88822
      lcronan
      Participant

      OK, I found a method that works:

      Hydroxylamine Mutagenesis. I got sooo many mutants, now it’s time for screening…

      I tried one protocol that asked for incubation at a high temp for a short amount of time. This didn’t make sense to me since hydroxylamine is denatured at high temps, but I tried it anyway and got no results. I tried another that asked for incubation at 37 for 20hrs and that seems to have worked beautifully!

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