Hello

That is tricky! Do you have standard curve? If you need to do compare expression of 2 genes in one cell population, reaction kinetics and efficiency must be the same. Otherwise your comparison wont tell you the truth!

Yes, but it would be semi quantitative as you wouldnt have a standard with an absolute known value to compare against.

Thank you all very much!

Since I only want to know which of the transcripts is more abundant, I think I will take Mohamed Darwish

I understand that my findings won't be very precise, but at the moment that's enough for me.

Thank you all again!

Its your decision anyway, but I don't think you need a reference gene in your case! Both of your genes came from the same sample! What could be the out come of normalization?

Ali Javadmanesh Thank you for your concern. The problem is that I have almost no experience in relative quantification of mRNAs, therefore, it's hard for me to imagine what could the outcome be. As you asked before - no, I do not have standard curve. I'm not sure if I can make one in my case - if you have any advice or protocol on it, I would gladly consider doing it this way

The best way of making an standard curve is to make a serial dilution of your cDNA. You may try 5 points (dilutions): Point 1= is the original cDNA, Point 2= 3 times diluted with ddH2O, Point 3= 3 times dilute from the second point, Point 4= 3 times dilute from the third point and Point 5= 3 times dilute the forth point. So you have 5 serial dilutions with different concentrations. From each concentration, you need to have a triplicate. The out come of these 15 reactions will provide you the standard curve. You will need standard curves for each of primers. If you have 2 gene, then 2 standard curves are needed. Volume of template used in each reaction should be equal to your unknown samples.

I wish I could be there to help you ;)