Updates are available Erratum dated 2018-02-28

Article History

Received: 28 November 2016

Accepted: 18 April 2017

First Online: 15 May 2017

Change Date: 28 February 2018

Change Type: Erratum

Change Details: Nat. Methods 14, 695–698 (2017); published online 15 May 2017; corrected after print 28 February 2018 We were alerted by readers that the reported Nm consensus sequence in mRNA matches the 3′-adaptor sequence used in sequencing library preparations, and this could be caused by mispriming1. In our approach, the majority of RNA fragments without Nm at the 3′ end are blocked from ligating to the 3′ adaptor because of the presence of 3′ phosphate from the last oxidation elimination step (OE) (Fig. 1a of the original paper), while Nm sites accumulate at the 3′ ends (Supplementary Figs. 1 and 2; Supplementary Notes 1 and 2). However, because of the low Nm abundance in messenger RNA (mRNA), only very limited amounts of mRNA fragments carry 3′ Nm and thus can be successfully ligated to the 3′ adaptor. Mispriming could occur if the 3′ end of the reverse transcription (RT) primer hybridizes to a few bases of the 5′-ligated RNA (Fig. 1a). Although our method effectively identifies Nm sites in abundant ribosomal RNA (rRNA, Supplementary Fig. 1), its application to less abundant mRNA can be contaminated by mispriming, leading to false-positive Nm sites and the erroneous AGAUC motif on mRNA (original Fig. 2d), which corresponds to the 5′-end sequence of the 3′ adaptor.

Competing interests

: The authors declare no competing financial interests.