Ligation question.

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    • #12725
      Darwin420
      Participant

      Hey guys,

      I am currently in a 3rd year molecular lab and I am conducting my first ligation next week.

      I already digested my fragment of interest however I made a silly error.

      For my ligation, I am also required to run a gel for the fragment of interest. I was suppose to set aside 1.5 microlitres of DNA with loading dye in a separate tube, where it will be ran on a gel. The remaining solution of DNA is the DNA I am going to ligate.

      HOWEVER, I am an idiot and put the loading dye to all my DNA, even the DNA I am suppose to ligate.

      Will the dye effect my ligation process?

      like intuitively I would say that it wouldn’t effect the ligation process… it doesn’t really react with DNA, that is why it is safe to add to DNA when we run it on a gel.

      However, has any one done this before? lol…

      what’s your insight? do you think my ligation will go fine?

    • #97344
      JackBean
      Participant

      Hi,

      you can use any of kits for DNA purification.Either some for purification after PCR or run electrophoresis with all the DNA and then cut it out and purify.
      Or use just phenol:chloroform, that could work too, but I don’t guarantee.

    • #97377
      Darwin420
      Participant

      Yea…but, what if I just added my solution with the dye and tried to ligate…do you think that would really mess up my ligation though?

    • #97378
      Darwin420
      Participant

      the reas why I ask this is because I don’t have the lab time to to purify the DNA from the loading dye.

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