Hyb & Seq™ NGS Technology

Simple, fast, scalable and accurate targeted sequencing

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The Hyb & Seq NGS Technology is being developed as a library-free, amplification-free, hybridization-based single molecule next-generation targeted sequencing workflow.

  • <30 minutes hands-on-time with sample to result in a single day
  • Simultaneous and direct DNA and RNA sequencing (without cDNA conversion)
  • Low and medium throughput sequencers in one system with standard flowcell (20M reads) and extended flowcell (120M reads)
  • Single-molecule reads with high fidelity (>99.9% accuracy), >99.99% (QV40) consensus accuracy and data free of systematic errors

Why Hyb & Seq technology?

Hyb & Seq technology offers several key advantages:

 

  • Simplest workflow to reduce hands-on time, labor costs, and required skill level
    No library prep or amplification eliminates error-prone sample preparation steps, creating a simple workflow that makes NGS easy for experts and beginners. You will be able to run NGS inhouse without the stresses of a complex and inefficient workflow, going from sample to sequencer with less than 30 minutes hands-on-time.

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  • Simultaneous DNA and RNA sequencing enables streamlined detection of all genomic variants
    Direct DNA and RNA sequencing without cDNA conversion results in a simple, fast, and more efficient workflow. Hyb & Seq provides the unique ability to detect all genomic variants (SNVs, CNVs, Indels, fusions) from a single tube and single workflow for an even more efficient and cost effective sequencing solution.

 

  • Flexible throughput to fit your needs
    The Hyb & Seq NGS system is optimized to provide cost-effective solutions for labs focused on targeted panel sequencing. Hyb & Seq has two flowcell options, a standard flowcell and an extended flowcell, which together provides affordable and flexible throughput options so your lab can scale from 200 to 2,500 samples per year per system.
Flowcell

Single molecule reads

Standard 20M
Extended 120M
  • Highly accurate single molecule sequencing
    Native single molecule sequencing with each base sequenced by multiple probes produces high fidelity reads (>99.9% QV30 accuracy and >99.99% QV40 consensus accuracy) without the systemic errors found in other technologies.

 

  • Less than 24 hours turnaround time
    Cyclic nucleic acid hybridization of sequencing probes directly onto native DNA and RNA targets enables sample to result in as little as 4 hours for pathogen identification and <24 hours for oncology panel sequencing.

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The Hyb & Seq technology enables direct, single-molecule sequencing of captured target sequences

 

Sample processing

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Prior to processing on the Hyb & Seq Instrument, target-specific capture probe pairs are hybridized to fragmented DNA or RNA molecules from genomic sample material. On the Hyb & Seq Instrument, these captured nucleic acid molecules, with a sequencing window of up to 100bp, undergo core sample processing which prepares the captured nucleic acid molecules for the subsequent sequencing steps. During the core sample processing, the captured targets are purified and pooled. Purified and pooled targets are then transferred to the sequencing card where they are hybridized to the flowcell's surface.

 

Sequencing

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Sequencing of these targets is accomplished through multiple sequencing cycles which involve cyclic nucleic acid hybridization of targets with sequencing probes, followed by readout with  reporter probes. Sequencing probes contain a hexamer sequencing domain and a reporter domain. The sequencing domain forms the complement to the target to be sequenced, and the reporter domain is a cyclically-read barcode. The reporter domain encodes the identity of the hexamer sequence hybridized to the target and is read via hybridization with fluorescently labeled reporter probes. A comprehensive set of sequencing probes with all possible hexamer sequences (4096) enable sequencing of any given target sequence. Hexamer sequences derived from each single target molecule are assembled using a graph-based algorithm and the resulting contiguous sequence reads are output into an industry-standard data output file (BAM or CRAM) that includes sequence quality metrics.

Hyb & Seq is a versatile sequencer capable of covering any NGS application. Targeted applications explored during technology development phase have included:

Oncology

  1. Clinically actionable, focused solid tumor cancer panel
  2. Targeted hematological oncology panel
  3. Comprehensive broad cancer panel

Infectious Disease

  1. Pathogen identification and resistance gene profiling
  2. Phenotypic antibiotic susceptibility test

Gene Expression Profiling

  1. Targeted RNA panels (10 - 5,000 genes)
  2. Transcriptome gene expression panel (22,000 genes)

For Research Use Only. Not for use in diagnostic procedures. 

The Hyb & Seq platform is currently in development targeting beta-instrument availability in 2019.