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) (primary sources 15–17) and of high-throughput sequencing technologies (possibly as high as 10^−2–10^−3 per nucleotide) (primary sources 18, 19) 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 (ref 20 BNID 113571) and 10^−12 (refs 21, 22 BNID 113571) per nucleotide, making it possible to measure error rates across the entire transcriptomes of viruses and other organisms." |