I think its wrong to say annealing only happens between 50-65 C, but
that's the general range for most primers 15-25 bp in length
On Fri, Jan 27, 2012 at 11:57 AM, veera <drveeramanikandan@gmail.com> wrote:
> generally annealing temperatures will be just 3-4 C below Tm of primers. if
> the melting point of primers is very high like you said in the range of 90 ,
> at that hight tempertaure all double stranded dna will split off and so
> annealing will not take place., because annealing happens only in the range
> of 50 - 65 C. hope i'm correct.. if i'm wrong anyone can correct me...
>
> Jonathan Nesser wrote:
>
>
> Sorry to start yet another new thread for such a small question, but I
> don't want to pollute other threads with off topic posts... I've been
> trying to understand why primer melting temperatures have to be in the
> 55-65C range when PCR cycles call for 94C for DNA melting... I've
> tried to find an explanation for this but haven't been able to...
> Wouldn't that result in the primers splitting off from the template
> DNA before you reach the extension cycle of 75C (or around there)? I'm
> sure I'm wrong because obviously these PCR programs work, but I don't
> know WHY I'm wrong, and would like to understand. Thanks again for
> explaining something that is probably so common knowledge that nobody
> feels the need to address it in writing :)
>
> Jonathan Nesser
> jonathan.nesser@gmail.com <mailto:jonathan.nesser@gmail.com>
> diybioandneurosci.blogspot.com <http://diybioandneurosci.blogspot.com>
>
>
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Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics
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