Taq activity?

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dzrdez
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Taq activity?

Post by dzrdez » Thu Aug 31, 2006 1:28 pm

What's the activity of Taq DNA polymerase at low temperature(20,30or 40 degree)?
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sdekivit
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Post by sdekivit » Thu Aug 31, 2006 6:38 pm

Taq polymerase is an enzyme with an optimum curve. So at lower temperatures as you mentioned Taq polymerase will get less active as far as I know.

--> It's use because the PCR reaction needs to take place at relatively higher temperatures than our body temperature. That's why you need Taq polymerase because it's resistant to higer temps.

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Post by oppox » Thu Aug 31, 2006 11:55 pm

Why are u planning on going so low?
I cant see why you shouldnt try it out, maybe u should increase your extansion time though, to 5-6 min perhaps.

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Post by oppox » Fri Sep 01, 2006 2:55 pm

on the other hand after some thoughts, in a ordinary PCR cycle the annealing temp is about 50 deg and then rised to 72 for taq to begin. So I dont know but it might be a problem there. If taq was active at so much lower temps, it would already have replicated the whole gene before the temp hit 72, am I fishing out of the blue now?

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Post by sdekivit » Fri Sep 01, 2006 4:20 pm

what would you expect to happen when you use normal DNA polymerase in a PCR and you let the temperature rise to 95 degrees C for 15 sec to separate the DNA strands ? There lies the choice of using Taq-polymerase: it's heat stable.

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