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qPCR with several BIOLOGICAL REPLICATES, how to compare?


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#1 AlessandraCastaldi

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Posted 18 October 2012 - 02:37 AM

Hello everybody,

I am performing a relative study with several controls (12 different individuals) and I want to put them in average in order to compare it with my Transgenic. Thus, I run a real-time PCR with all my controls and all my samples on the same plate. My question is: how can I do the average of controls? Should I do the calculation of DDCt and then do the average of Fold Changes, or could I average the DCt of controls (DCtav) and then calculate DDCt based on the DCTav for my samples?

I hope you can help me!
Thanks
Alessandra

#2 Trof

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Posted 20 October 2012 - 01:15 PM

Usual way is to mix all control samples together equally, and use this pooled sample as a calibrator in DDCt calculations.
It's also necessary to run all control samples too, individually to see the range of controls compared to the range of transfgenics.

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