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  • Visualizing phosphorothioate oligonucleotides

    I would like to somehow visualize phosphorothioate oligonucleotides or use another tool to ascertain whether adding S-Oligos into an active cell cultures allows for entry and possibly integration into the genome.

    From this article, it appears that someone has successfully done so in bacteria, but they introduced a mutated oligonucleotide that integrated into the bacterial genome, and the mutation conferred some apparent phenotype.

    Click here for article

    My experiment should be innocuous in terms of phenotype, but I still need a way to know if it entered and integrated.

    Thanks!
    Your BRO.

  • #2
    How is this next gen sequencing or de novo
    discovery?

    Comment


    • #3
      Sorry ECO I am new to this forum and didn't know the appropriate place to put the thread. If this is an error I humbly apologize and would appreciate if you could tell me the correct discussion location!

      Comment


      • #4
        Is there a sequencing approach here? I cant understand how your question relates to sequencing.

        Moving to General.

        Comment


        • #5
          what do you mean by visualize? Modeling by a MD simulation? Or actually see them in the cell? My guess is attach some kind of fluorescent labels but don't know how you can directly visualize the phosphorothioate backbone. Yeah, and, this topic is totally not NGS related, I think.

          Comment


          • #6
            I am probably not using the terminology right. By "visualizing" I meant, "confirm" that my oligonucleotide entered and integrated into the cells. The sequence itself already exists in the cell, but I am adding s-oligos (phosphorothioate, basically where they replace one of the oxygens on the linking phospho with a sulfur) and need to confirm their entry/integration. I don't necessarily need to see them. Again, sorry for posting this, looks like I'm not even in the right forum for this kind of question. Do you know where I can get help?

            I have looked into fluorescent labels but the problem as I understand it is that fluorescent label groups are bulky and would interfere with integration in active culture, mitosis etc. Its fine for FISH and things like that but not for what I am planning.

            -Your BRO.

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