Im trying to design an experiment at the moment to look at the difference in expression of three genes between trained and untrained athletes.
Im not looking for copy number just expression of the gene relative to a housekeeper.
So for example, after the experiments are run i can state that relative to beta-actin the untrained athletes express 4 times less of GENE A than the trained athletes.
I am able to get a blood sample from each of the participants and extract genomic DNA from the blood.
Now i am attempting to design my QPCR experiment.
1.Can i use genomic DNA in my QPCR (Syber green) reaction?
2.When designing the primers do i need to design them within an exon to avoid really long intronic sequences?
3.Is there any recommended housekeeping genes used when dealing with gDNA?
ideally i would like to be able to get RNA from the blood and make cDNA, however it is often 24/48 hours after it being drawn before i receive the blood.
Any ideas or comments would be gratefully received.