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Single Molecule, Real-Time Technology
Revolutionize Genomics
with SMRT® Sequencing
Resolve to Master Complexity
The complex genomes of plants and animals, with their
multi-gigabase sizes, polyploidy, and dificult-to-sequence
repetitive regions, hold the key to resolving agricultural and
environmental challenges like drought and disease. With
a complete view of genomes and transcriptomes of crops,
livestock, and associated microbes, scientists can inally
unlock the genetic diversity required to advance breeding,
precision engineer genes, develop novel treatments and
natural growth enhancers, and secure a global food supply.
Infectious diseases are responsible for more than 23% of
global deaths, including 50% of child mortality. Antibiotic
drug resistance is a major threat to global health security,
extending far beyond the human health sector, and
globalization has created vast opportunities for novel
diseases to emerge, spread, and kill. Only comprehensive
characterization of these pathogens including their mobile
elements will lead to the discovery and design of better
vaccines, treatments, and outcomes.
Despite large investments in population studies, the
heritability of the majority of Mendelian and complex
diseases remains unclear, limiting development of
diagnostics and treatments. Shedding light on the complete spectrum of sequence variant types with
chromosome-level phasing across genomes unique to a
population, disease or individual may provide a holistic
view of human genetics to resolve missing heritability
linkages.
The most comprehensive view of
genomes, transcriptomes, and epigenomes
Sequel® System
Accelerate Your Science
pacb.com/sequel
A SMRT Foundation
Single Molecule, Real-Time (SMRT®) technology is built upon two key innovations
that overcome major challenges in the ield of sequencing. Zero-Mode Waveguides
(ZMWs) allow light to illuminate only the bottom of a well in which a DNA
polymerase/template complex is immobilized. Phospholinked nucleotides allow
observation of the immobilized complex as the DNA polymerase produces a
completely natural DNA strand.
SMRT Cells containing up to a million ZMWs are processed on PacBio® Systems
which simultaneously monitor each of the waveguides in real time.
Zero-Mode Waveguides
Primer
TemplatePolymerase
Phospholinked Nucleotides
Up to a million ZMWs per SMRT Cell
SMRTSequencing
Advantages
High
Accuracy
Uniform
Coverage
Epigenetics
Single-
Molecule
Resolution
Long
Reads
SMRT Sequencing Achieves
Long Read Lengths
Read length data shown above from a 35 kb size-selected human library using the SMRTbell® Express Template Prep Kit on a
Sequel System (3.0 Chemistry, Sequel System Software v6.0, 20-hour movie). Read lengths, reads/data per SMRT Cell and other
sequencing performance results vary based on sample quality/type and insert size among other factors.
Re
ads
Read Length
Half of data in reads: >45 kbData per SMRT Cell: Up to 20 Gb
0
50,000
100,000
150,000
200,000
250,000
300,000
0 50,000 100,000 150,000 200,000 250,000
Half of data in reads:
>45 kb
Top 5% of reads:
>150 kb
Longest read lengths:
>200 kb
Re
ads
High-Fidelity, Long Reads
Long reads with Q20 single-molecule accuracy
Estimated number of high-idelity, long reads (Q20 or Q30) per Sequel SMRT Cell (Chemistry 3.0. Sequel System Software v6.0)
based on insert size and movie collection time.
0
100,000
200,000
300,000
400,000
500,000
600,000
1000 (10 hr) 2000 (10 hr) 5000 (20 hr) 10000 (20 hr)
Q20 Q30
High Consensus Accuracy
Consensus accuracy is a function of coverage and chemistry. The data above is based on a bacterial genome run on the
Sequel System (3.0 Chemistry, Sequel System Software v6.0) Single-molecule accuracy has similar coverage requirements.
Free of systematic errorsAchieves >99.999% (QV50)
PACIFIC BIOSCIENCES® CONFIDENTIAL
10.0
20.0
30.0
40.0
50.0
60.0
70.0
0 20 40 60 80 100 120
Uniform Coverage
Se
qu
en
ce
cove
rag
e
QV
(S
ing
le o
r M
ulti-
mo
lecule
)
Percent GC contentCoverage
No ampliication requiredEven coverage across GC content
Mean coverage per GC window across a human sample.
60
50
40
30
20
0
0 10 20 30 40 50 60 70 80 90 100
Num
be
r of
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Insert Size (Movie Time)
Comprehensive Genomics
pacb.com/publications
Complete Knowledge
• Affordably generate gold-standardmicrobial genomes
• Detect and resolve plasmids, mobileelements, and structural variationincluding gene duplication and inversion
• Simultaneously analyze genome-widemethylation with single-base resolution
Complete genome assembly and methylome (red spikes) of an E. coli strain with six plasmids (not to scale).
Unobstructed Views
• Sequence low-complexity regions,like trinucleotide repeats
• Access all variant types, includingstructural variants, Indels and SNVs
• Allele-speciic phasing ofhaplotypes in targeted regions orbetween chromosomes
Haplotype 1
Haplotype 2
PacBio coverage
heterozygous 280 bp deletion Structural Variants
A heterozygous deletion structural variant downstream of the gene TMEM2 is
GRCh38 chr9 71,654,000 71,655,000 71,656,000
A heterozygous deletion structural variant downstream of the gene TMEM2 is supported by half of the PacBio reads that map to the locus. Sequence data is from the human sample HG002.
Conident Discoveries
• Directly detect full-length transcriptswithout assembly
• Characterize gene-isoform expressionwithin targeted genes, or across anentire transcriptome
Full length isoform sequences from brain tissue of Anna’s hummingbird (red transcript models) identify two additional non-coding 5’ exons
(purple arrows and inset) and extend 3’ UTRs (green arrow) while also capturing all ive known splice variants (blue transcript models).
For Research Use Only. Not for use in diagnostic procedures. © Copyright 2018, Paciic Biosciences of California, Inc. All rights reserved. Information in this document is subject to change without notice. Paciic Biosciences 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 Paciic Biosciences products and/or third party products. Please refer to the applicable Paciic Biosciences Terms and Conditions of Sale and to the applicable license terms at http://www.pacb.com/legal-and-trademarks/terms-and-conditions-of-sale/.
Paciic Biosciences, the Paciic Biosciences logo, PacBio, SMRT, SMRTbell, Iso-Seq, and Sequel are trademarks of Paciic Biosciences. BluePippin and SageELF are trademarks of Sage Science. NGS-go and NGSengine are trademarks of GenDx. FEMTO Pulse and Fragment Analyzer are trademarks of Advanced Analytical Technologies. All other trademarks are the sole property of their respective owners.
Flexible Design and Analytics
• Flexible run time less than a day
• Serially process up to 12 SMRT Cellsin a single run
• Walk away time up to 4 days
• Size-selection options toenrich for longest inserts
• Multiplexing and barcodingsolutions available
• Variety of analysis methods available throughSMRT Link and PacBio DevNet community
• Open source software
• Advanced data visualization and mining
Comprehensive de novo assemblies
Target all types of variants across relevant genomic regions
Full-length isoform transcripts
Resolution of complex populations
Methylation pro�les
• Express template preparation in as few as 3 hrs
• Accepts a variety of sample types and insert sizes
Sequel System
www.pacb.com
Headquarters
1305 O’Brien DriveMenlo Park, CA 94025 United StatesPhone: 1.650.521.8000
Singapore Oice
20 Science Park Road #01-22 TeleTech Park Singapore 117674Phone: 65.6778.5627
Customer Service
Phone: 1.877.920.PACB (7222) option 1Fax: 1.650.618.2699Email: [email protected]
Technical Support
Phone: 1.877.920.PACB (7222) option 2Email: [email protected]
PacBio® Sequencing Providers
www.pacb.com/SMRTproviders
Inquiries
North America: [email protected]
South America: [email protected]
Europe/Middle East/Africa: [email protected]
Asia Paciic: [email protected]
PN: BR108-100318
