quantifying spice variant
I am interested in analyzing a gene that has two splice variants. I know that both of them are expressed - as seen by semi-quantitative RT-PCR. I can design unique primers only against 1 of the two variants. Primers targeting the second variant also amplify the first one (though the amplicon sizes are different).
I want to investigate if there is any difference in their temporal profiles or not. I assume that I can do that by looking at the melt curve data (I should get two peaks since the product size is different – by ~100bp).
But is there any way I can quantify their expression levels?
Another related question – how to quantify changes in their expression levels under different conditions?
I have never worked with genes having multiple splice variants, so I’ll appreciate any help regarding this issue. Thank you very much.