Abstract
The PSeq platform begins with preparing DNA libraries with a specialized informational architecture. Conventional NGS paired-end sequencing then provides the automated data pipeline with all the information required to assemble each individual RNA. Together, these elements form PSeq Clear-Signal Architecture™, an integrated chemistry-to-data framework for whole molecule sequencing. This paper presents initial technical validation for PSeq v1 across the entire chain of operations.
In a case study, data assembled for a single transcript are readily visualized with an open-source gene viewer (IGV) designed for examining whole-genome sequences and RNA-Seq data.
Stepping through this information for a single transcript from the gene PRPF31 visualizes the pipeline performance at a level of detail not otherwise available. This exercise further illustrates the traceability of all steps in the platform chain of operation.
This white paper constitutes initial technical validation, not to be construed as a statistically complete platform validation, clinical validation or regulatory performance analysis.
Key topics
- Bioanalyzer library-size profiles and marker-position evidence
- Sanger confirmation of marker, barcode, and check-base structure
- Representative FASTQ tag-plus-cDNA structure and same-gene read pairing
- PRPF31 SMID-bin evidence for single-gene confinement and splice-junction phasing
- Raw-read-to-consensus traceability and current validation limits
Contents
- Library-validation questions and evidence summary
- Library size distribution and marker placement
- Sanger confirmation and FASTQ read composition
- PRPF31 molecule-level case study
- Exon alignment and end-to-end splice-junction coverage
- Shotgun consensus assembly and base-level error traceability
- Interpretation of current evidence and planned next-phase validation