[Bioperl-l] Bio::SeqFeature::Primer Calculating the Primer TM
redwards at utmem.edu
Mon Feb 16 22:40:40 EST 2004
I wrote this implementation of the Bio::SeqFeature::Primer module, and
I am guilty as charged. I was confused by the primer3 documentation,
but hey, at least I was honest in the docs - I said that I couldn't
figure out what was going on.
I don't know what the formula is that Primer3 used. If you have some
code for this I can update the module.
On Feb 12, 2004, at 5:50 AM, john herbert wrote:
> Hello BioPerl.
> I was thinking of using BioPerl to calculate the Tm of my primers. I
> looked from the documentation and found the method for getting a
> primer's Tm. my $tm = $primer->Tm;
> In the notes of this method, the author is confused by the fact that a
> BioPerl calculated Tm never matches that of Primer3's prediction. This
> is because Primer3 uses a different method to calculate a Primers TM
> than the method used in the BioPerl.
> BioPerl $primer->Tm = Calculated using: Tm = 81.5 + 16.6(log10([Na+]))
> + .41*(%GC) - 600/length
> (Sambrook, Fritsch and Maniatis, Molecular Cloning, p 11.46 (1989, CSHL
> Primer3 primer TM = Primer3 uses the oligo melting temperature formula
> given in Rychlik, Spencer and Rhoads, Nucleic Acids Research, vol 18,
> num 12, pp 6409-6412 and Breslauer, Frank, Bloeker and Marky, Proc.
> Natl. Acad. Sci. USA, vol 83, pp 3746-3750. This method uses
> thermodynamics to calc TM.
> Primer3 does use the Sambrook method to predict the TM of the PCR
> products but not the Primer TM.
> Would it be possible to update the BioPerl to match how Primer3
> calculates the Primer TM?
> Kind regards,
> John Herbert
> Cancer Research UK.
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> Bioperl-l at portal.open-bio.org
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