Post on 06-Oct-2020
transcript
Introducing HiSeqTM 2000HiSeq 2000
Abizar Lakdawalla
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Abizar Lakdawallaalakdawalla@illumina.com
Simple Illumina Sequencing Workflow
SIMPLIFIED SAMPLE PREP
cBot CLUSTER GENERATION
HiSeq 2000 SEQUENCING
DATA PROCESSING & ANALYSIS
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Parallel sample processing
Automated clustergeneration
Automated sequencing
Simple, efficient data analysis
HiSeq 2000 Comparison with the Genome Analyzer
HiSeq 2000(at launch)
GAIIx
(at 50G)
Gb per run 150-200 50
GAIIx
(at 95G)
95
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Gb per runGb per dayCluster density in KClusters/mm²**Read lengthAvailable surface area (mm²)*
150-20020-25
260-3502 x1002880
505
4902 x100
510
*GAIIx with single surface, single FC, HiSeq 2000 with dual surface, dual FC**Clusters passing filter
957
6202 x150
510
HiSeq 2000 - two surface scanning
1. Line scanner2. Epi-illumination3. Top surface (all 4 colors) scanned in one pass4. Bottom surface (all 4 colors) scanned in 2nd pass4. Bottom surface (all 4 colors) scanned in 2nd pass5. 2 scanning swaths in each lane of a flow cell
Clusters
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Bottom surface
Top surface
HiSeq 2000New flow cell design
LARGER, DUAL-SURFACE ENABLED
>5x increase in imaging area>5x increase in imaging area
Retains 8 lane format
Compatible with cBot
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HiSeq 2000 Dual Flow Cell DesignExperimental flexibility
SIMPLE FLOW CELL LOADING
Flow cells held by vacuum
No oil needed
LED switch ensures correct connection
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HiSeq 2000 Plug-and-play reagents
PRE-CONFIGURED SEQUENCING REAGENTSPRE-CONFIGURED SEQUENCING REAGENTS
Only two minutes hands-on time
Up to 200 cycles per flow cell
Bar-coded for tracking
Temperature-controlled compartment
Integrated paired-end fluidics
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HiSeq 2000A leap forward in sequencing innovation
MOST WIDELY ADOPTED AND PROVEN CHEMISTRY
Leverages Illumina’s reversible terminator based SBS chemistry Leverages Illumina’s reversible terminator based SBS chemistry
INNOVATIVE ENGINEERING
Dual flow cell, dual surface imaging
Four camera system with cutting edge scanning technology
HUMAN INTERACTION DESIGN ELEMENTS
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HUMAN INTERACTION DESIGN ELEMENTS
Simplest user experience
HiSeq Control SoftwareTouch screen user interface
SIMPLE OPERATION
Step-by-step run set up
Real time run monitoring
Reagents usage tracking
Remote monitoring from web browser
Enter read length, single, paired-read, indexing
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Remote monitoring from web browser
Multimedia help menu
HiSeq Control SoftwareTouch screen user interface
SIMPLE OPERATION
Step-by-step run set up
Real time run monitoring
Reagents usage tracking
Enter read length, single, paired-read, indexing
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HiSeq Control SoftwareTouch screen user interface
SIMPLE OPERATION
Step-by-step run set up
Real time run monitoring
Reagents usage tracking
Remote monitoring from web browser
Enter read length, single, paired-read, indexing
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Remote monitoring from web browser
Multimedia help menu
HiSeq 2000 Data AnalysisA simple, familiar workflow
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HiSeq CONTROL SOFTWARE
Read Generation
CASAVA
Alignments, variations, builds
VISUALIZATION
GenomeStudio, or favorite browser
HiSeq 2000Sequencing redefined
HIGHEST OUTPUT
200 Gb per run at launch200 Gb per run at launch
FASTEST DATA RATE
~25 Gb/day
7-8 days for 2 x 100 bp
HIGHEST NUMBER OF READS
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HIGHEST NUMBER OF READS
One billion single-end reads
Two billion paired-end reads
Whole Transcriptome Studies: Two Runs
INDIVIDUAL HUMAN TISSUES
16 tissues, 1 lane per tissue, 2x50 bp
Standard poly A+ mRNA-Seq Library Preps
Body Map 2.0 ProjectAdrenal, Adipose, Brain, Breast, Colon, Heart, Kidney, Liver, Lung, Lymph Node, Ovary, Prostate, Skeletal Muscle, Testis, Thyroid, White Blood Cells
TOTAL HUMAN TRANSCRIPTOME
Equal Mixture of 16 Human Tissues
3 libraries–polyA+ mRNA, normalized polyA+, normalized ribominus RNA method
Strand-specific mRNA-Seq, 1x100 bp
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Format PF readsAverage error rates
(phiX spike-ins) Yield
Individual human tissues 2x50 2.24 B 0.27%, 0.18% >100 Gb in <5 days
Total human transcriptome 1X100 1.19 B 0.67%, 0.81% >100 Gb in <5 days
Thyroid, White Blood Cells
SUMMARY OF RUN METRICS
HiSeq 2000Unmatched cost-effectiveness
HiSeq 2000$690K
cBot$55K
$690K
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Total run cost US List
Price per run (2 flow cells, 2 x 100 bp, 200 Gb)
Price per human genome (1 flow cell, 100 Gb)
$19,800
$9,900
*available shortly after launch
Consumables cost estimates
Bacterial genomes (5Mb at 30x depth)
– 1000 – 2000 samples per run
– $10-20 + sample prep cost / sample
Whole human exomes (30Mb at 50x depth)
– 100-200 samples per run
– $100-200 + sample prep cost / sample
Human transcriptomes (20M templates/sample)
– 50 samples per run (50bp)
– $200 + sample prep cost/sample
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– $200 + sample prep cost/sample
Whole human genome (3Gb at 30x depth)
– 2 samples per run
– $10000 per sample
Thank you
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