show Abstracthide AbstractWe devised a novel normalization approach employing chimpanzee RNA as a spike-in standard in NGS-based human gene expression analysis. In this approach, human transcripts were normalized read-by-read by spiked-in chimpanzee transcripts that were readily discriminated by natural single nucleotide variations (SNVs) between the two species. The SNV-normalized analysis enabled more accurate measurement of differential gene expression than an ordinary read-based approach.