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Accuracy of real-time PCR results?

Accuracy of real-time PCR results? - Protocols and Methods Forum

Accuracy of real-time PCR results? - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 09-25-2003, 02:08 PM
Paul Wary
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Default Accuracy of real-time PCR results?



What accuracy can I expect from real-time PCR results (in percent of
relative n-fold differences)?

Paul
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Old 09-25-2003, 02:29 PM
Duncan Clark
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Default Accuracy of real-time PCR results?

Historians believe that in newspost
<[Only registered users see links. ]> on Thu, 25 Sep 2003,
Paul Wary <[Only registered users see links. ]> penned the following literary
masterpiece:

It's probably more dependant upon how good your pipetting of
template/dilutions of template is, than anything else.

You can definitely determine a 2 fold difference in copy no i.e. 10,000
and 5000 copies - 1 Ct apart. I would not like to do the same for 20 and
10 copies.

To tell the difference between 10000 and 7500 as opposed to 10000 and
6000, I would never do a call on. Below one Ct you are getting a bit
'subjective' given that multiple replicates may be 0.25Ct's apart
(getting worse as copy no. decreases).

Duncan
--
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.
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