Revio Long-read Sequencing System Performance Enhancements

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Overview

Since its introduction, Pacbio SMRT sequencing technology has gained attention for its advantages of long reads (10-25k), single molecules, and no GC preference. In 2019, PacBio launched HiFi sequencing, which increases single-molecule sequencing accuracy to 99.9% (Q33). And now, the next-generation, high-throughput Revio system, launched in 2022, breaks through the throughput limit once again. With advanced sequencing consumables, robotics, and powerful computational capabilities, Revio delivers accurate long reads and methylation information directly from the sequencer in every run. Unleash yourself to discover more with a complete view of the genome, epigenome, and transcriptome on the Revio System. The Revio System provides metagenomics researchers with faster, more cost-effective long-read sequencing, resulting in higher-quality and more complete microbial datasets and access to new areas of discovery.

What is the Revio System?

Revio is a gene sequencing system for human genetic analysis, cancer research, agricultural genomics, etc. The Revio System supports HiFi sequencing. It provides the foundation for long read, superior accuracy, and direct methylation detection, adding affordability, high throughput, and ease of use.

Advantages of the Revio System

  • Higher Throughput Driven by Workflow Efficiency
    The Revio System utilizes high-density SMRT Cell chips with 25 million ZMWs each, capable of running up to 4 SMRT CELL chips in parallel, with a 24-hour runtime yielding 360 Gb of HiFi reads per day, which is the equivalent of sequencing 1,300 entire human genomes per year.
  • Complete Phased Human Genomes for as Little as $1,000
    HiFi sequencing provides small variants, structural variants, repeat amplification, methylation, and haplotype profiling of the human genome in a single library build and sequencing run.
  • Simplified Reagent Consumables and Flexible Instrumentation
    Compared to the Sequel IIe system, the Revio system cuts consumable usage in half and offers significant workflow and convenience improvements: the newly designed SMRT Cell eliminates the need for nitrogen supply to protect DNA, allows for simultaneous setup of subsequent sample runs while the current sample is being sequenced, and allows for timely replenishment of sequencing consumables.
  • 20X Increase in Computational Power & Improved HiFi Yield and Accuracy
    The Revio system has built-in NVIDIA GPUs to process all data in real-time without the need for any offline analysis. PacBio has also established a partnership with Google Health to integrate DeepConsensus+ deep consensus algorithms into the system, which improves HiFi yield and accuracy. Additionally, the Revio system reduces file size per base by 50%, which is only 1/20th of the file size of other long read technologies.

Exclusive Customized Workflow

The consumables and software used with the Revio System are flexible and easy to use, greatly simplifying the steps and optimizing the experience. Whether you’re in a genome center or a small lab, you’ll experience it all:

Fast and reliable library preparation

Library construction for the Revio System supports manual or automated preparation using the SMRTbell Preparation Kit 3.0.

Fast run setup

Sequencing plates and run designs are automatically connected via an NFC tag, allowing run setup on the instrument in less than a minute.

Highly adaptable to your experimental schedule

The Revio system works independently of the four microarray benches, so the system can be loaded with consumables for up to 20 hours per day, ensuring full utilization of each microarray bench.

Powerful computing

Built with multiple powerful NVIDIA GPUs, the Revio System delivers up to 20 times the computational power of the Sequel IIe System, delivering instantly available and accurate data directly from the instrument.

HiFi read generation

GPUs reduce turnaround time for base detection and HiFi read generation, outputting HiFi reads directly in BAM format.

Google Health DeepConsensus

DeepConsensus uses advanced deep learning techniques to further improve the accuracy and throughput of HiFi sequencing.

Methylation detection

Standard runs include DNA methylation status, calculated using deep learning algorithms capable of handling polymerase kinetics.

Splitting

All basic processing steps, including barcode splitting, are done on the instrument.

Enhanced HiFi Sequencing with the Revio System

Preserve the benefits of HiFi sequencing Get more information on the Revio system
– Provides accurate long read of natural DNA molecules
– Use of Nanobind® CBB kits and SMR
– CBB kit and SMRTbell® preparation kit 3.0.
– Easy sample and library preparation
– Generate HiFi reads + methylation detection, barcode splitting on the instrument
– Standard BAM file format compatible with downstream analysis tools
– High throughput – up to 1,300 HiFi genomes per year
– Onboard Google DeepConsensus for the most accurate HiFi reads available
– Simpler user experience with fewer consumables for fast runs
Setup
– 50% smaller output files for reduced storage costs

Diverse Applications for the Revio System

Whole Genome Sequencing

The Revio System excels at whole-genome sequencing, providing de novo assembly and variant detection. Its ability to sequence tens of bases in HiFi reads allows researchers to easily resolve large variants and navigate challenging genomic regions.

Targeted Sequencing

For more specific research requirements, the Revio system offers targeted sequencing. Whether it’s sequencing amplicons that can process more than 1,000 samples in a single run, or targeted enrichment across the 20 Mb to 100 kb range, Revio meets a wide range of research needs.

Metagenomics

The Revio system brings significant advantages to metagenomic research. Whether analyzing 384 communities through birdshot metagenomics or assembling data from 12 communities, the depth, and accuracy of the Revio system bring clarity to complex microbial datasets.

RNA Sequencing

For transcriptomics, the Revio System provides single-cell transcriptomics tools that ensure comprehensive data of over 80 million reads per library.

Reference

  1. Manuel, Juana G., et al. “High Coverage Highly Accurate Long-Read Sequencing of a Mouse Neuronal Cell Line Using the PacBio Revio Sequencer.” bioRxiv (2023): 2023-06.

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Written by kikogarcia