Quantitation methods

a_firzinger at my-deja.com a_firzinger at my-deja.com
Mon Feb 21 05:55:11 EST 2000


In article <88hg3f$mdj$1 at newsg2.svr.pol.co.uk>,
  "Bucky" <buckminster at fullerene.freeserve.co.uk> wrote:

Another method for quantification would be "Online" or
"real-time" PCR. You would need a TaqMan from PE or a
Light Cycler.
For competitive PCR, which is as far as I know always
only valid for semiquantification, the dilution of your
competitive standard is critical (you should no exactly
how many copies of your standard you have in your PCR).
Another method could be determining the number of cycles
the PCR needs to leave the exponential phase.
(e.g. Gonzalez, P., J. S. J. Zigler, et al. (1999). “Identification and
isolation of differentially expressed genes from very small tissue
samples.” Biotechniques 26(5): 884-6, 888-92.)

If you want to determine the number of integrations
into your transfected cell, then I would suggest inverse
PCR. How would you have done this by competitive PCR?

Regards,

Andreas :-)


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