Please I have been working on PCR for some days now but on quant studio analysis, it has been showing no amplification. I have tried repeating the protocol with several dilutions of cDNA(1:5, 1:10, 1:20) but no change in results. I attempted gene expression on two different housekeeping genes(ACTB &HPRT1) on the pig's ileum but no amplification was seen on analysis.
I might be useful to know if the pcr is failing or if it is working but the quant studio setting are not set correctly so you are not picking up the increased signal of amplification. Try running a couple of your amplified samples on an agarose gel to see if you have any amplification. While you are doing that if any of your colleagues have primers for any housekeeping gene borrow some and run a pcr for gel running to test if your cdna is good quality and also as a check that your primers are/are not working