| Comments |
P.3311 right column top paragraph: "RNAseq offers an approach to both disentangle transcription errors from translation errors and provide an error rate for every transcribed gene in a genome. Unfortunately, the high error rates both of cDNA synthesis (3–6 × 10^−5 per nucleotide) (refs 15–17 BNID 113568) and of high-throughput sequencing technologies (possibly as high as 10^−2–10^−3 per nucleotide) (refs 18, 19 BNID 113568) renders the transcription errors obtained by conventional RNAseq indistinguishable from sequencing artifacts. Two recently developed methods offer ways to circumvent these problems by allowing transcription errors to be distinguished from sequencing and cDNA synthesis errors. Through the use of altered library preparation protocols, these methods reduce the overall error rate of RNAseq to less than 10^−8 (primary source 20) and 10^−12 (primary sources 21, 22) per nucleotide, making it possible to measure error rates across the entire transcriptomes of viruses and other organisms." |