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 ? |
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| 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 ? |
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| 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 ? |
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| No, dilution series do not need to be repeated. |