Revio system

Reveal more with accurate
long-read sequencing at scale

Scale

Ease of use

Compute power

Affordability

2,500 human HiFi
genomes per year

Only 500 ng DNA input
and flexible run setup

Google DeepConsensus
and more on board~$500 human
HiFi genome

This is your moment for a more complete view of biology

More comprehensive variant
calling with phasing and 5mC+ 6mA methylation detectionMore complete
assembly ofcomplex genomes

Targeted sequencing
to study genes
at scaleSingle-celltranscriptome
sequencing

Access high accuracy for all
variant types — SNVs, indels,
structural variants, tandem
repeat expansions, andmethylation — including inchallenging regions.1

Accurately construct
the full sequenceof chromosomes,including telomeres andcentromeres.2

Focus the power of HiFi
variant calling by enrichingfor regions of interest using
PureTarget™, hybrid capture,or PCR amplification.

Catalog RNA isoforms at
single-cell resolution, moving
beyond gene counting
to catalog full-lengthtranscripts.3

See it all with HiFi sequencing on the Revio system
Long reads

Exceptional accuracy

HiFi reads are tens of kilobases long, providing the ability toresolve large variants — like structural variants and tandem
repeat expansions — and map to difficult, repetitive regionsof the genome.

HiFi sequencing provides reliable answers throughexceptional accuracy, with 90% of bases ≥Q30 and medianread accuracy ≥Q30.

Direct methylation detection in native molecules

Low DNA input requirements

HiFi sequencing identifies base pair-level DNA methylation in
all runs, without any additional library prep steps.

Each sample requires only 500 ng of DNA input, enabling
HiFi sequencing for even the most precious samples on theRevio® system.

Uniform coverage across sequence contexts
With an optimized polymerase and no amplification, HiFi
sequencing provides uniform coverage even for high-GC
regions and tandem repeats.
84 kb

CFTR
HiFi reads for the sample HG002 reveal accurate sequence of both maternal and paternalalleles, providing phasing that spans over 84 kb of the cystic fibrosis gene, CFTR.

Ignite your research with SPRQ chemistryWith SPRQ™ chemistry, the Revio system helps you achieve more with less DNA, increasing efficiency, maximizing output, andreducing costs—empowering you to make more discoveries faster and affordably.

Spark innovation:• Resolve chromatin architecture with Fiber-seq, poweredby on-instrument 5mC and 6mA calling, now with anincreased 5mC CpG accuracy for even greater precision

Reduced input, expanded application possibilities:• Only 500 ng DNA input• 4× more efficient SMRT® Cell loading

Drive down genome costs:• 33% increased HiFi yields• 2 human genomes at 20× per SMRT Cell

Multiomics in every run
Independent stages enable simultaneous sequencing of 4 SMRT Cells, delivering high-quality, haplotype-resolved data for thegenome, transcriptome, and epigenome – all from a single run.
SMRT Cell 1

SMRT Cell 2

SMRT Cell 3

SMRT Cell 4

2 whole humangenome at 20×

384 microbialgenomes

6 full-length transcriptomes
at 10M reads

8 human methylation
profiling at 5×

PacBio Revio with SPRQ

SBS short-read sequencing

Nanopore sequencing

Read length

15–20 kb

2×150 bp

10–100 kb

Read accuracy

99.95% (Q33)

99.92% (Q31)

99.26% (Q21)

24 hours

44 hours

72 hours

100–120 Gb

2,400–3,000 Gb

50–110 Gb

Variant calling—SNVs

Variant calling—indels

Variant calling—SVs

5mC, 6mA

5mC, 6mA

How Revio compares

Run timeYieldMethylationPhasing

The easy-to-use Revio system fits your schedule
The Revio system offers configurable
sequencing run times to tailor to yourexperiments.
The new flowcell design of the Revio SMRT
Cell eliminates the need for a nitrogen supply,
simplifying laboratory requirements.

Available
to load

The ability to load consumables while
sequencing is in progress makes it easy
to maximize instrument utilization without
off-hours work.

12

3

4

Independent
sequencing stages

The workdeck on the Revio system is isolated
from the four sequencing stages. This leaves
the system available for loading consumables
up to 20 hours per day while keeping thestages fully utilized.

Enjoy a fully automated sequencing workflow

4
independent

sequencing stages

1–4
SMRT Cells

24
hrrun times
Simplified consumables• The Revio system has only three workdeck consumables— a sequencing plate, a SMRT Cell tray, and pipette tips.

• Now optimized with SPRQ chemistry, the Revio system
adds a low DNA input requirement, enhanced affordability,and even higher throughput to a foundation of long reads,• Sequencing plates are linked automatically to run designs
exceptional accuracy, direct methylation detection, andthrough an NFC tag, enabling rapid run setup on instrument.
uniform coverage• The Revio SPRQ sequencing plate combines what waspreviously four parts, eliminating extraneous plastic andmaking runs easier to manage. The single standard 96-wellplate includes sample libraries, reagents, and space formixing and waste.

Get the most accurate reads — fast• The GPUs provide rapid turnaround time for basecallingand HiFi read generation, keeping pace with thesequencing throughput of the Revio system.
• With Google DeepConsensus4 on board, Revio is the mostaccurate PacBio long-read system to date. DeepConsensus
uses advanced deep learning techniques to further extend
the accuracy and yield of HiFi sequencing.

HIFI READS• The Revio system features cutting-edge NVIDIA GPUs with>20× more compute power than the Sequel® IIe system.

Q20

Q30

Q40

READ ACCURACYSequel® IIe

DeepConsensus on Revio

Reduce your compute and storage costs• The Revio system is capable of much more than justbasecalling. Every run also measures DNA methylation
status, calculated with a deep learning algorithm thatprocesses polymerase kinetics.

10.0
bytes/base• All fundamental processing steps are performed on
instrument, including barcode demultiplexing andconversion to the standard BAM format.
• A more efficient data representation reduces file size by
50% per base — 20× smaller than for other long-readtechnologies. Base quality scores are grouped into sevenbins, and similar reads are sorted together for more
effective compression.

1.0
bytes/basePrevious format

0.5
bytes/base
Revio BAM

On instrument

Off instrument• Base calling• Variant calling• HiFi read generation with DeepConsensus• Genome assembly• Methylation calling

Revio system• Barcode demultiplexing• BAM file generation• Signal collection• Additional base calling• Initial base calling• Methylation calling• Barcode demultiplexing

Other long-readtechnology• BAM file generation• Variant calling• Genome assembly

fast5/pod5

Add to what you love about HiFi sequencing with the Revio system
Keep what’s great about HiFi sequencing…and get even more on the Revio system• Long, accurate reads of native DNA molecules• High-throughput — up to 2,500 HiFi genomes per year

• Easy sample and library prep with options for manualand automated workflows• 500 ng of DNA input per sample• On-instrument generation of HiFi reads + methylation
calls (5mC and 6mA), with demultiplexing• Standard BAM file format compatible with downstream
analysis tools• A simplified user experience, including adaptive loading,fewer consumables, rapid run setup, and real-time
performance run previews• Onboard Google DeepConsensus for the most accurate
HiFi reads yet• Reduce storage costs with smaller output files at only
~60 Gb each

350K

500K
450K

300K

400K

250K

HIFI READS

300K
250K
200K
150K
100K

0K

200K
150K
100K
50K

50K
0

5K

10K

15K

20K

25K

30K

0K

35K

Q20

Q30

Q40

Q50

READ QUALITY

READ LENGTH, BP

97.7 99.8 99.9

80

92.1

98.0 99.1

93.2

96.7 97.2

70

ACCURACY, F1%

60

BASES %HIFI READS

350K

5040302010x20x30x

100Q0Q10Q20Q30

BASE QUALITY

Q40SNVSINDELS

Data shown is for a single Revio SMRT Cell for HG002/GM24385 sequenced with SPRQ chemistry.5SVS

System specifications with SPRQ chemistry
The Revio system utilizes nanofabricated Revio SMRT Cells and has four independent stages, allowing sequencing of multiple
SMRT Cells in parallel.
Library

Run time*

Q30+ bases

HiFi yield per SMRT Cell†

1–5 kb

12 hours

95%

6–8 million reads

5–10 kb
10–15 kb

35–70 Gb
24 hours90%

15–20 kb
20–25 kb

Methylation

5mC at CpG sites
and 6mA ‡ fornative DNA

70–100 Gb
100–120 Gb

30 hours

85%

100–120 Gb* Run time refers to the data collection step, which determines the time between processing SMRT Cells.
†HiFi yield is dependent on library quality and sequencing preparation procedures. Specified yield is based on high-quality samples prepared following best practices.

‡The Revio 6mA caller is designed to detect methylation in the context of the Fiber-seq chromatin assay.6

Key applications and sample throughput with SPRQ chemistry
The Revio system supports a variety of applications that benefit from accurate long HiFi reads and requires only 500 ng ofinput DNA per sample.
LibraryApplication

Per Revio SMRT Cell

Per year*

Expected coverage†

1–5 kb

Amplicon sequencing>1,000>2.5 million

50×

5–10 kb

Microbial genome

384

480,000

30×

5–10 kb

PureTarget repeat
expansion panel

48

60,000200×15–20 kb

Human genome

22,50020×

15–20 kb

Human methylation
profiling8

10,0005×

15–20 kb

Transcriptome withKinnex™ full-length
RNA kit

67,500

10 million reads* Annual throughput is based on 2,500 Revio SMRT Cells for 12 hour runs; 1,250 for 24 hour runs; and 1,050 for 30 hour runs. Adaptive loading — which increases theconsistency of yield per SMRT Cell — adds about 4 hours to run time, affecting the maximum number of SMRT Cells per year.
†Expected coverages are estimates.

What will you discover with Revio?
Learn more about the Revio system:

We have offices in countries around the world.
Visit pacb.com/contact for contact info.
Headquarters
1305 O’Brien DriveMenlo Park, CA 94025
United States
Phone: 1.650.521.8000Customer service
1.877.920.PACB (7222), option 1
Fax: 1.650.618.2699
orders@pacb.com
Technical support
1.877.920.PACB (7222), option 2
support@pacb.com
Connect with PacBio for more info:
North America: nasales@pacb.com
South America: sasales@pacb.com
EMEA: emea@pacb.com
Asia Pacific: apsales@pacb.com

pacb.com/revio
Ordering information
Part number

Product102-090-600

Revio system103-260-000

Nanobind PanDNA kit

102-182-700

SMRTbell prep kit 3.0

103-520-100

Revio SPRQ polymerase kit

102-202-200

Revio SMRT Cell tray

103-504-900

Revio SPRQ sequencing plate

Instrument operating environment
Temperature

19–25°C (66–77°F)

Humidity

20–80% relative humidity, non-condensing

Ventilation

13,000 BTU (3,800 W)

W×D×H

92.7 × 91.4 × 174.5 cm (36.5 × 36.0 × 68.7 in)

Weight465 kg (1,025 lb)

Power

200–240 VAC at 50–60 Hz, 30A

Network

1 GbE or 10 GbE, copper

Noise output<75 dB at 3.3 m (10 ft 10 in) from instrument

KEY REFERENCES
1. Lincoln, S. E., et al. (2021). One in seven pathogenic variants can be challenging to detect by NGS: an analysis of 450,000 patients with implications for clinical sensitivity and genetic test
implementation. Genetics in Medicine, 23(9), 1673–1680.
2. Nurk, S., et al. (2022). The complete sequence of a human genome. Science, 376(6588), 44–53.
3. Al’Khafaji, A. M., et al. (2021). High-throughput RNA isoform sequencing using programmable cDNA concatenation. bioRxiv, doi:10.1101/2021.10.01.462818.
4. Baid, G., et al. (2022). DeepConsensus improves the accuracy of sequences with a gap-aware sequence transformer. Nature Biotechnology, 1–7.
5. PacBio (2022). HG002 sequence data from Revio system.
6. Stergachis, A., et al. (2022). Single-molecule regulatory architectures captured by chromatin fiber sequencing. Science 368, 1449-1454.
$500 per human HiFi 20X genome assumes US list price of $995 for sequencing reagents for one Revio SMRT Cell, which has an expected yield of 120 Gb, equivalent to two 20x human genomes.
Research use only. Not for use in diagnostic procedures. © 2024 Pacific Biosciences of California, Inc. (“PacBio”). All rights reserved. Information in this document is subject to change without notice. PacBio
assumes no responsibility for any errors or omissions in this document. Certain notices, terms, conditions and/or use restrictions may pertain to your use of PacBio products and/or third-party products. Refer tothe applicable PacBio terms and conditions of sale and to the applicable license terms at pacb.com/license. Pacific Biosciences, the PacBio logo, PacBio, Circulomics, Omniome, SMRT, SMRTbell, Iso-Seq, Sequel,Nanobind, SBB, Revio, Onso, Apton, Kinnex, PureTarget, SPRQ, and Vega are trademarks of PacBio.

102-326-543 REV06 31OCT2024