the target per copy of the nontarget portion of the genome. For single-copy genes, N will have a value of 1 and can be ignored. However, in the case of multicopy targets such as mtDNA or ribosomal genes, N can be on the order of hundreds or thousands of copies per nontarget genome and can markedly influence the amount of input DNA. The value of N can be reasonably estimated by performing copy number analysis with quantitative PCR (qPCR) using primers that are specific to the target and nontarget DNA. Similar to the nontarget DNA inputs, the target DNA input amounts should also be proportionally increased with increasing lane fraction (Table 2).
