Has anyone here referred to a housekeeping gene to normalize their target gene data? I have recently done some experiments where I tried to normalize my control and treated samples with rpl-32 and actin. Both times, I could see this variability in Ct values across samples with housekeeping gene primers. Could anyone share their experience regarding this? I am really confused as to what is going on since I was expecting Ct values for rpl-32/actin to be very similar across samples.
There is a set of housekeeping genes to choose from and with the geNorm program you can choose the most stable one for your samples… It is recommended that you use more than one housekeeping gene for normalisation.