I have some samples that have not gone through PCR yet. I need to know the ideal PCR cycle because I am targeting a yield of a certain concentration and quality of sample. I don't have enough sample volume to test PCR at different cycles. I use SYBR to measure everything (qPCR, Qubit). I have a qPCR kit with SYBR green mastermix and DNA standards. How do I use this kit to set up qPCR and determine the number of cycles and quantity at that cycle? For example, I want the qPCR to show that if I performed PCR on my samples, then at 14 cycles, I will get 500nM of product. Now when I do perform PCR on the samples at 14 cycles, I will indeed get 500nM or around there.
Does anyone have a protocol to set up something like this? I have qPCR SYBR DNA standards, SYBR green qPCR mastermix, and qPCR primers needed to qPCR amplify my samples.