qPCR with several BIOLOGICAL REPLICATES, how to compare? - (Oct/18/2012 )
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!
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.