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which of the following is NOT TRUE with regard to replication cycles used in

which of the following is NOT TRUE with regard to replication cycles used in - PCR - Polymerase Chain Reaction Forum

which of the following is NOT TRUE with regard to replication cycles used in - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 06-03-2011, 08:18 PM
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Default which of the following is NOT TRUE with regard to replication cycles used in



Polymerase Chain Reaction? HERE 'C' STANDS FOR CELSIUS

A. 90-96C - DNA is denatured into single strands
B. 50-60C - The primers anneal to their complementary sequences
C. 72C - The polymerase binds and extends a complementary DNA
strand from each primer
D. 80C - The polymerase adds dNTp's from 3'- 5, reading the
template from 5'- 3'
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Old 06-03-2011, 08:18 PM
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D.

The polymerase has to attach the dNTP to a free OH group at the 3' end. This means that the dNTP's are added in a 5' > 3' direction.
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Old 06-04-2011, 05:49 AM
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Default Re: which of the following is NOT TRUE with regard to replication cycles used in

uhm... extension of PCR amplicon generally optimum at 74C. Yes most of us use 72C I believe it is to prolong the life of the Taq. At 80C its activities is lowered. I don't know how to say this...

A: 90-96 does denature the DNA to single stranded but also depend on the duration.
B: yes, if the template still in single stranded mode.
C: Hmmmm
D: Correct would be direction 5' - 3' and 3' - 5' were to reversed. [The polymerase adds dNTp's from 5'- 3', reading the template from 3'- 5']

Yes, Maxorz say D is same with mine but my reason is at 80C they don't really extend that much... most time will be start going thru disassociate. If not all PCR protocol will want to stay longer in 80C rather than 72C right? and the above too.
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