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The right temperature for unfolding RNA secondary structure

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The right temperature for unfolding RNA secondary structure

Postby frankalpha » Wed Sep 22, 2010 9:10 pm

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.
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Postby JackBean » Thu Sep 23, 2010 8:20 am

I think that also depends on the secondary structure ;) But the limit is probably due to the enzyme
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Re: The right temperature for unfolding RNA secondary structure

Postby frankalpha » Thu Sep 23, 2010 2:33 pm

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?
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Postby trimethoprim » Fri Sep 24, 2010 1:43 am

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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