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Real Time PCR polymerase enzyme

real time PCR

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3 replies to this topic

#1 Mehdi

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Posted 11 March 2012 - 05:04 AM

hi guys


A typical TaqMan protocol contains a denaturation step followed by a combined annealing and extension step at 55–60°C, instead of the traditional three-step PCR cycle of denaturation, annealing, and extension. my question is, does combined temperature affects Taq polymerase efficiency? and which Taq polymerase we use for Taqman Real Time assay?

#2 Trof

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Posted 12 March 2012 - 04:41 AM

Is it a homework question?
What types of Taq polymerase do you know if you're saying "which Taq"?

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#3 tea-test

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Posted 12 March 2012 - 04:59 AM

At least, it should have a 5' to 3' exonuclease activity for TaqMan assays. Posted Image

The lower temperature will affect the TaqPol, the velocity of extension will be decreased (this is one reason why you should keep real-time PCR products short).
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#4 Mehdi

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Posted 12 March 2012 - 09:07 AM

At least, it should have a 5' to 3' exonuclease activity for TaqMan assays. Posted Image

The lower temperature will affect the TaqPol, the velocity of extension will be decreased (this is one reason why you should keep real-time PCR products short).


Thank you so much. your post answered my question.

Edited by Mehdi, 12 March 2012 - 09:08 AM.






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