Assessing PCVR inihibitors in a ABI 7700 SDS

Yves Bertheau yves.bertheau at free.fr
Sun Feb 27 02:55:06 EST 2000


Hello,

We would like t appreciate the quality of several methods to extract
DNA. We would like to use  real-time PCR in an ABI 7700 SDS to
assess the presence of, at least, soluble PC inhibitors.

The first trials showed that adding foreign DNA to an internal PCR
control do not change the Ct value, as expected, but the "slope" of
the exponetial phase and the end-point values (amplicons yield
values).... 

Thus what can we use to measure the effects of the PCR inhibitors?

Do you know 

1. which kind of appropriate measure, ready to use or not from the ABI
7700 SDS machine, we could use?
2. in case of negative answers how to recover data to establish or
already calculated  "slope" of the exponential phase?
2. in case of negative answers how to recover the data for the end of
the exponential phase, (or the beginning of the plateau effect)? 
If possible not the raw data but after subtraction of the
background....

Thanks for you rhelp

Regards

Yves Bertheau
INRA PMDV / MDO
Route de Saint CYR
78026 VERSAILLES cedex
bertheau at versailles.inra.fr




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