Normalizing seems like a lot of trouble. Is it really necessary?
YES!!! It’s a couple of hours spent at the beginning when you are in a hurry to get results, but it’s many hours saved later (not to mention money) when you are trying to analyze data, and some of it is off scale and some has dropped out, and then you must cherry pick your way through the plate and redo the PCR on those that didn’t work.
