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. 2019 Nov 22;10:5292. doi: 10.1038/s41467-019-13228-9

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — The principle and validation of PAIso−seq. a Flowchart for PAIso−seq method design. The main steps of the method include end-extension, template-switching, full-length cDNA amplification, circular adaptor ligation, and PacBio sequencing. b The structure (top panel) and agarose gel analysis of the poly(A) spike-ins (middle panel). The mean poly(A) tail length of each spike-in measured by PAIso−seq (bottom panel). Source data are provided as a Source Data file. c The poly(A) tail lengths of Dnmt1, Btg4, and Plat in GV oocytes measured by PAIso−seq (density plot of poly(A) tail length of detected CCS reads of the given genes, three replicates, left panel) and PAT assay by using capillary electrophoresis on fragment analyzer (mean of three replicates, right panel). The mean length of each gene poly(A) tail measured by PAIso−seq is shown. The number of CCS reads used are 141 (Dnmt1/GV rep.1), 249 (Dnmt1/GV rep.2), and 165 (Dnmt1/SCGV com.); 164 (Btg4/GV rep.1), 521 (Btg4/GV rep.2), and 357 (Btg4/SCGV com.); 136 (Plat/GV rep.1), 277 (Plat/GV rep.2), and 207 (Plat/SCGV com.). The average length of poly(A) tails are 74 nt (Dnmt1), 44 nt (Btg4), and 45 nt (Plat) measured by the PAT assay. RFU, relative fluorescence units.