A Simplified Workflow: Democratizing Study of Isoforms using the Sequel System

Full length RNA isoform sequencing using Single Molecule, Real-Time (SMRT) sequencing (Iso-Seq analysis), has often resulted in numerous novel discoveries and insights into RNA processing and function. Indeed, studies have... [ view full abstract ]

Full length RNA isoform sequencing using Single Molecule, Real-Time (SMRT) sequencing (Iso-Seq analysis), has often resulted in numerous novel discoveries and insights into RNA processing and function. Indeed, studies have shown unique discoveries even on samples once considered having well characterized transcriptomes. It is obvious from these outcomes that the ability to sequence full length RNA isoforms will yield new biological understandings. However, challenges have remained that impede routine adoption of the method.

Here, we will present a new breakthrough on the Sequel System in the ability to load a range of RNA transcripts (1 kb – 6kb+) in a single SMRT Cell. This enhancement has resulted in a simplified workflow with only one library required – compared to four before – to achieve complete transcriptome coverage. Further, it reduces hands-on steps and has minimized the need for size selecting a library thus reducing time to results. We will also describe new enhancements to SMRT Analysis’ Iso-Seq protocol which now provides an optional mapping to a reference genome post clustering to obtain the final set of unique transcript isoforms and updates to our TOFU algorithm.