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Common analysis of direct RNA sequencinG CUrrently leads to misidentification of m 5 C at GCU motifsRNA modifications, such as methylation, can be detected with Oxford Nanopore Technologies direct RNA sequencing. One commonly used tool for detecting 5-methylcytosine (m5C) modifications is Tombo, which uses an “Alternative Model” to detect putative modifications from a single sample. We examined direct RNA sequencing data from diverse taxa including viruses, bacteria, fungi, and animals. The algorithm consistently identified a m5C at the central position of a GCU motif. However, it also identified a m5C in the same motif in fully unmodified in vitro transcribed RNA, suggesting that this is a frequent false prediction. In the absence of further validation, several published predictions of m5C in a GCU context should be reconsidered, including those from human coronavirus and human cerebral organoid samples.more » « less
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Dunning Hotopp, Julie C.; Baltrus, David A.; Bruno, Vincent M.; Dennehy, John J.; Gill, Steven R.; Maresca, Julia A.; Matthijnssens, Jelle; Newton, Irene L.; Putonti, Catherine; Rasko, David A.; et al (, Microbiology Resource Announcements)null (Ed.)ABSTRACT Microbiology Resource Announcements (MRA) provides peer-reviewed announcements of scientific resources for the microbial research community. We describe the best practices for writing an announcement that ensures that these publications are truly useful resources. Adhering to these best practices can lead to successful publication without the need for extensive revisions.more » « less
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