PCR optimization

Susanne Rohrer "srohrer(removethis)" at immv.unizh.ch
Thu Aug 3 04:06:56 EST 2000



Tina wrote:

> Does the predetermined optimal cycle number for a set of primers (being
> within the linear portion of the amplification curve) change among
> different blood donnors?
> (Note : I work on human leukocytes)

Since the amount of template affects the amount of product that is
amplified, if you have more, then you will be out of the linear range
quicker.

It depends on how great the variation is in the number of leukocytes a
person has in his/her blood (leukemia? CD4+ in HIV?).

It also depends on how uniformly you process your samples.

To answer such questions to yourself, I think you really have to visualize
the process of PCR, and you will understand it. Same with one of your
previous questions about primer orientation.






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