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NAD-seq for profiling the NAD+ capped transcriptome of Arabidopsis thaliana

ABSTRACT: Summary Eukaryotic RNAs can be modified with a non-canonical 5′ nicotinamide adenine dinucleotide (NAD+) cap. NAD-seq identifies transcriptome-wide NAD+ capped RNAs. NAD-seq takes advantage of click chemistry to allow the capture of NAD+ capped RNAs. Unlike other approaches, NAD-seq does not require DNA synthesis on beads, but this technique uses full NAD+ capped transcripts eluted from beads as the substrates for strand-specific RNA sequencing library preparation. For complete details on the use and execution of this protocol, please refer to Yu et al. (2021). Graphical abstract Highlights • Protocol for NAD-seq allowing eukaryotic NAD+ capped transcript discovery• Details all steps necessary for producing NAD-seq libraries• Instructions for analysis workflow to identify eukaryotic NAD+ capped transcripts Eukaryotic RNAs can be modified with a non-canonical 5′ nicotinamide adenine dinucleotide (NAD+) cap. NAD-seq identifies transcriptome-wide NAD+ capped RNAs. NAD-seq takes advantage of click chemistry to allow the capture of NAD+ capped RNAs. Unlike other approaches, NAD-seq does not require DNA synthesis on beads, but this technique utilizes full NAD+ capped transcripts eluted from beads as the substrates for strand-specific RNA sequencing library preparation.

SUBMITTER: Yu X

PROVIDER: S-EPMC8593656 | biostudies-literature |

REPOSITORIES: biostudies-literature

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NAD+-capped RNAs are widespread in the Arabidopsis transcriptome and can probably be translated.

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