Fig. 5: Characterization of SCAN-seq and analysis of unannotated transcripts.
From: hnRNPM cooperates with BCAS2 to modulate alternative splicing during oocyte development
a Schematic illustration of the SCAN-seq method. Briefly, control and Hnrnpm cKO GV oocytes were lysed to release mRNAs. First-strand cDNAs were synthesized using a 24-nt barcode RT primer. Following amplification, the cDNAs derived from multiple cells were combined, pooled together, and purified. Subsequently, the full-length cDNAs were used for the construction of nanopore libraries and sequencing. The mouse, oocyte, and nucleic acid schematics are modified adaptations of original resources from SciDraw and Servier Medical Art, respectively, and are employed under a CC BY 4.0 license. b–j PCR and Sanger sequencing validation of genes (Nlrp14, Meikin, Vrk1, and Zar1l) with novel isoforms identified in GV oocytes. Ref. Reference. Representative results shown in (b–e) were obtained from at least three independent experiments with similar results. Source data (b–e) are provided as a Source data file.
