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Primer concentrations with rapid PCR

Robert Hartley r.hartley at freeuk.com
Mon Jan 3 20:25:54 EST 2000

In message <3870fdbe.5193211 at news.gwdg.de>
          nvahsen at hot[nospam]mail.com    (remove the [nospam]) (Nicolas von Ahsen) wrote:

> Hi Lars,
> I think you are right.
> For a 20mer primer at 0.2 M Na-equivalents you can calculate a t1/2 of
> approx. 3 sec. for annealing. And annealing time is should be short on
> the LightCycler. More than 10 sec. makes no sense.

10 sec annealing? 1sec if using capiliaries. 20 with tubes.
 Also I use a standard mix in my RapidCycler.9no optics)
I beleive the increase is in Mg since the borosilicate adsorbs a lot of Mg.

Contact DNAamp for their mix. I have never raised the primer conc with my 
machine using their stuff. I may change this now.

> Read Wetmur JG, Crit Rev Biochem Mol Biol 1991, 26:227 for more.

I will check out this.
> Best regards
> Nicolas

Now more and more are using this technology does anyone know of a forum or a 
mail list for these machines. Light cycler and Rapid from Idaho technologies

Bob Running an Intel StrongARM PC600 with RiscOS 3.7 

Robert Hartley, Partick, Glasgow. Home of the RiscOS Molecular Biology page 
Home  r.hartley at freeuk.com    http://home.freeuk.com/r.hartley 
Work  rh at mblab.gla.ac.uk      http://www.gla.ac.uk/centres/cellengineering/

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