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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
>

cheers
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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