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http://schema.org/contentSize 8266 https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/contentUrl https://api.rohub.org/api/ros/3543b082-9077-492e-a4c7-a3b7c8bb39e8/crate/download/ https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/creator mailto:wltsangac@connect.ust.hk https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/dateCreated 2025-07-29 00:00:00 https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/dateModified 2025-10-16 11:11:58.719757+00:00 https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/datePublished 2025-07-29 00:00:00 https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/description Polyadenylation is a dynamic process which is important in cellular physiology. Oxford Nanopore Technologies direct RNA-sequencing provides a strategy for sequencing the full-length RNA molecule and analysis of the transcriptome and epi-transcriptome. There are currently several tools available for poly(A) tail-length estimation, including well-established tools such as tailfindr and nanopolish, as well as two more recent deep learning models: Dorado and BoostNano. However, there has been limited benchmarking of the accuracy of these tools against gold-standard datasets. In this paper we evaluate four poly(A) estimation tools using synthetic RNA standards (Sequins), which have known poly(A) tail-lengths and provide a valuable approach to measuring the accuracy of poly(A) tail-length estimation. All four tools generate mean tail-length estimates which lie within 12% of the correct value. Overall, Dorado is recommended as the preferred approach due to its relatively fast run times, low coefficient of variation and ease of use with integration with base-calling. https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/encodingFormat application/ld+json https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/exifData #accessions https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/exifData #additionalInfo1 https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/exifData #additionalInfo2 https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/exifData #additionalInfo3 https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/exifData #additionalInfo4 https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ http://schema.org/exifData #additionalInfo5 https://w3id.org/ro-id/3543b082-9077-492e-a4c7-a3b7c8bb39e8/ 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Reames B. 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