Exercise 7: Dilution Series for Calculating Amplification Efficiencies

TODO Convert this to h5p

In my qPCR experiment I want to study 8 new genes for which I had to design new primer pairs in 12 samples (samples of interest and control samples). I want to use 2 technical replicates and 96 well plates.

What is the first thing I need to do ?
Perform a pilot experiment to determine the amplification efficiencies of these primer pairs.

For this I need a dilution series of representative cDNA template.

How many dilutions would you include ?
A dilution series with 6 dilutions for 8 genes nicely fits into a 96 well plate.

A few weeks after my initial qPCR experiment I want to test these 8 genes in a new set of samples.

Do I have to repeat the pilot experiment ?
No, dilution series do not need to be repeated.