Efficient detection of alternative spliced human proteome using translatome sequencing

Alternative splicing (AS) isoforms create numerous proteoforms, expanding the complexity of the genome. Highly similar sequences, incomplete reference databases and the insufficient sequence coverage of mass spectrometry limit the identification of AS proteoforms.In this work, we compared RNC-seq and Ribo-seq in the context of proteome identification, especially when identifying protein isoforms from AS. We also demonstrated that the single-molecule long read sequencing technique identified thousands of new splice variants and guided the MS identifications of new protein isoforms. Overall design: Getting translatome data of MHCC97H RNC and RPF.And we performed the direct full-length RNC-seq on MHCC97H using Nanopore MinION sequencer