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    • #13807
      frankalpha
      Participant

      As the title goes, what’s the right temp. for that? In RT using superscript II or III, we use 65°C. Can we go any higher and will this damage the RNA? Thanks.

    • #101441
      JackBean
      Participant

      I think that also depends on the secondary structure 😉 But the limit is probably due to the enzyme

    • #101444
      frankalpha
      Participant

      Sure. Say if I don’t put the enzyme in at first, just heat the RNA to unfold the secondary structure, what temperature should I use to best unfold the secondary structure without damaging the RNA? And if the temperature drops down (I will have to add enzyme anyway), will the structure re-form?

    • #101453
      trimethoprim
      Participant

      I think the RNA will re-fold when the temp drops, but the folding may not be the same as the original one as that folding is a random process outside cells.

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