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The Agilent Technologies
SureSelect™ Platform for Target Enrichment
Focus your next-gen sequencing on DNA that matters
Emily M. Leproust, Ph.D.
Director Applications and Chemistry R&D – Genomics
Agenda:
• Introduction
• Applications for Mendelian diseases research
• Custom biomarker discovery and profiling
• Applications for cancer biomarker discovery and profiling
• Automation
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 2
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 3
Target Enrichment: A Highly Enabling Process
What?
• Also referred to as genome partitioning, targeted re-sequencing, DNA capture…
• Captures genomic material of interest for next generation sequencer (i.e. Illumina, SOLiD, 454 etc…)
Why?
• Sequence your regions of interest!
• Enables focus on a subset of the genome
• Saves both time and money for downstream sequencing
• Identify homozygous and heterozygous variants
in targets relative to the reference genome
gDNA
Enriched library
Sequencing Technology Keeping up with Science
(from http://www.sciencemag.org/cgi/content/full/291/5507/1221/F1)
Older Methods
Appropriate
Lower Throughput
Discovery
PCR + Sanger
Sequencing
Newer Methods
Appropriate
High Throughput
Discovery
SureSelect™
Platform +
Next-gen
Sequencing
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing Workflow
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 5
Agilent’s SureSelect™ Platform: New Options
SureSelect Target
Enrichment System*Developed in collaboration
with the Broad Institute
Dr. Chad Nusbaum et al.
SureSelect
DNA Capture ArrayDeveloped in collaboration
with Cold Spring Harbor
Dr. Greg Hannon et al.
*Flagship Method
Agilent 60mer Array
1-3 µg gDNA
1-5 µg gDNA
(with WGA)
20 µg gDNA
(unamplified)
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 6
Distinct Target Enrichment Products for
Distinct Project Needs
SureSelect Target
Enrichment
System
SureSelect DNA
Capture Array
Throughput High Low
Study Sizes 10-1,000s samples 1-10 samples
DNA Input 1-3 µg* 1-20 µg**
Capture of
Target DNA
Up to 6.8 Mb (1x tiling, 120mer
baits) for custom; catalog products
1Mb (20x tiling)
(Custom 60-mer
baits)*minimum of 500 ng of prepped sample required
**Depends upon use of whole genome amplification.
or
0.5µg
<3µg
Library Preparation• Illumina• SOLiD
Hybridization / Capture24 hours
Baits:
- cRNA probes
- Long (120bp)
- Biotin labeled
- User-defined (eArray)
- SurePrint synthesis
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 7
Bead Separation
Wash / Elution / Amp
Advantages of Agilent Target Enrichment:
• Long baits tolerate mismatches
• RNA-DNA hybrids stronger than DNA-DNA
• RNA probe is strand-specific:
Allows large molar excess of bait,
Target-limited; improves uniformity
• Easily automated: All steps liquid handling
• Low input DNA (< 3 ug)
• 24hrs hybridization
• Same high quality Nucleic Acids as Agilent arrays
• Working on Solution Enrichment since 2006, license from Broad
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 8
SureSelect™ Target Enrichment System: Workflow
24 hours
0.5µg
<3µg
SOLiD 3
SOLiD 4
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 9
SureSelect™ Target Enrichment System: Workflow
Illumina GAIIx
HiSeq 2000
24 hours
0.5µg
<3µg
Page 10
Broad Paper on Cover of February, 2009 Nature Biotechnology Underlying Technology of SureSelect™ Target Enrichment System
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing Workflow
Agilent SureSelect™ Target Enrichment Efficacy:
Sample Coverage Plot
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 11
This plot (generated in the UCSC Genome Browser) shows sequence coverage data for a representative portion of the Agilent SureSelect Target Enrichment demo kit, which covers the RefSeq exons on the non-pseudoautosomal portion of Chromosome X using 2x tiling. In red is the log of the sequence coverage. Bait coordinates are denoted in black and the exons that constitute the target region are in blue. Sequence coverage is higher for those exons that are covered by more than one overlapping bait
269
0
SureSelect Bait Coordinates: Black Bars
2x tiling design approach
Agenda:
• Introduction
• Applications for Mendelian diseases research
• Custom biomarker discovery and profiling
• Applications for cancer biomarker discovery and profiling
• Automation
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 12
Exon capture is powerful to study Mendelian diseases
• Mendelian diseases are caused by coding mutations (with some exceptions)
• Exons are only ~1-1.4 % of human genome (30-50Mb)
• Primarily protein coding regions
Advantages:
• Much less sequencing
• ~5% of WGS, so up to 20x more samples
Disadvantage:
• Miss non-coding variants
Why coding+?
• More interpretable
• Easier to follow up
• Especially adapted to study of Mendelian diseases
• CCDS exons – v1
• CCDS + RefSeq – 38Mb v2 (Broad)
• GENCODE – 50Mb (Sanger)
• Includes ncRNA
• All Exons on X chromosomes
• 7674 exons
• 3Mb
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 13
Applications to Mendelian disorders
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 14
… and many more to come
• The Problem: Linkage analysis confirmed family inheritance pattern to a 28 Mb region (build 36)
• >200 genes contained in 28 Mb region; daunting task to sequence and validate all the variants.
• Solution: Identify a 2nd family that manifests the syndrome, determine linkage and perform targeted sequencing using the X-demo kit on heterozygote females of both families
Page 15
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing Workflow
• Target : 2.7 Mb; 36 bp SE seq on Illumina GA
• Findings
• Family 1 heterozygote showed a frameshift due to an A insertion in RBM 10 (RNA binding motif 10)
• Family 2 heterozygote showed a G to A substitution in RBM 10 (RNA binding motif 10)
• Both mutations were confirmed by Sanger Sequencing
• Inhibition of the orthologous Rbm 10 expression in developing mouse embryo using anti-sense probes showed patterns that were consistent with observed human malformations
Page 16
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing Workflow
Page 17
Human All Exon Kits – Most comprehensive coverage
Original design Exome V2 50Mb design
CCDS Sept. 2008
CCDS Sept. 2008 + additional RefSeq
content including CCDS Sept. 2009 exons
GENCODE and Sanger (includes CCDS and
Broad defined v2 content as well)
CCDS (Sept. 2009) 93.76% 99.01% 99.86%
CNV (Mar. 2010) 23.98% 27.49% 30.62%
Ensembl (6/16/2010) 65.58% 71.37% 75.24%
miRNA (miRBase 14) 90.00% 90.00% 92.78%
GenBank (6/16/2010) 75.96% 89.07% 90.74%
RefSeq Genes (6/16/2010) 86.69% 93.29% 96.47%
RefSeq Transcripts (6/16/2010) 88.85% 95.07% 97.50%
Total 37Mb 38Mb 50Mb
Developped with Broad Broad Sanger
• Human All Exon kits can be customized (PLUS) with up to 6.8Mb additional custom content • Human All Exon kits can be multiplexed on SOLiD4 and HiSeq2000
New
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing Workflow
Human All Exon 50Mb – 2x76bp, 50-60M HQ reads
Page 18
76.3%
85.1%
96.7%87.9%
77.5%
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
% on target +/-
100bp
Uniformity (3/4 mean with upper
tail):
% bases with 1x
coverage
% bases with 10x coverage
% bases with 20x coverage
Most comprehensive Human All Exon content available
38Mb design = a subset of 50Mb
Sequencing capacity: • 0.5-1 sample / lane GAIIx• 1-3 samples / lane HiSeq• 5-10 samples /flowcell SOLiD4
Chemistry recommended:• PE 2x76bp Illumina• PE 50+25 SOLiD
Multiplexing:• Illumina• SOLiD
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing Workflow
Comparison of SNP calls with HapMap
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 19
99.1% 99.2%98.4% 98.0%
95.7%94.9%
70%
75%
80%
85%
90%
95%
100%
Human All Exon v2 Human All Exon 50Mb
GT is REF GT is variant HOM
GT is variant HET
99.8% 99.7%98.2% 98.1%98.5% 98.3%
99.4% 99.3%
70%
75%
80%
85%
90%
95%
100%
Human All Exon v2 Human All Exon 50Mb
GT is REF GT is variant HOM
GT is variant HET OVERALL
Genotype Concordance vs. HapMapGenotype Sensitivity vs. HapMap
All Exon Plus (Add custom content to your All Exon)
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 20
All Exon Library
+
Your Custom Library
Enter Your Custom Regions in eArray
CCDS exons>1000 ncRNAs
38Mb
Your regions of interest
Is the Human All Exon not hitting all of your regions of interest?
Human All Exon Plus (All Exon + custom content)
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 21
1 tube capture, 1 lane seq. at 2x76bp on GAIIx = ~2Gb
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Exome +
0.87 Mb
Exome +
1.7 Mb
Exome +
3.4 Mb
Exome +
6.8 Mb
Exome
Control
SNP Analysis vs. HapMap
Sensitivity Concordance
0
5000
10000
15000
20000
25000
30000
35000
Exome +
0.87 Mb
Exome +
1.7 Mb
Exome +
3.4 Mb
Exome +
6.8 Mb
Exome
Control
22394
23224
24337
26352
21976
4480
5528
5816 6
182
5173
No. S
NP
s
SNP Analysis vs. dbSNP
Concordant Novel Mismatched
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 22
Efficient Capture of 5bp deletion on the
X-Chromosome: Menke’s Syndrome
hg18_ChrX_77131408_77131467_+ : Wildtype Bait Design
CTATTGTTTATCAACCTCATCTTATCTCAGTAGAGGAAATGAAAAAGCAGATTGAAGCT
CTATTGTTTATCAACCTCATCTT-----AGTAGAGGAAATGAAAA
ATTGTTTATCAACCTCATCTT-----AGTAGAGGAAATGAAAAAG
TTGTTTATCAACCTCATCTT-----AGTAGAGGAAATGAAAAAGC
GTTTATCAACCTCATCTT-----AGTAGAGGAAATGAAAAAGCAG
TATCAACCTCATCTT-----AGTAGAGGAAATGAAAAAGCAGATT
ATCAACCTCATCTT-----AGTAGAGGAAATGAAAAAGCAGATTG
ATCAACCTCATCTT-----AGTAGAGGAAATGAAAAAGCAGATTG
ATCAACCTCATCTT-----AGTAGAGGAAATGAAAAAGCAGATTG
CAACCTCATCTT-----AGTAGAGGAAATGAAAAAGCAGATTGAA
CCTCATCTT-----A-TAGAGGAAATGAAAAAGCAGATTGAAGCT
SureSelect™ Target Enrichment Kit Efficiently Captures 5 bp Mutant
Readout on Illumina GA
Agenda:
• Introduction
• Applications for Mendelian diseases research
• Custom biomarker discovery and profiling
• Applications for cancer biomarker discovery and profiling
• Automation
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 23
Other Applications of Targeted Re-Sequencing
• Capture any custom genomic regions (introns, exons, UTRs, regulatory, etc.)
• Ideal for biomarkers discovery and profiling (e.g. cancer)
• Ideal for custom SNP follow-up
• Ideal for characterization of large sample cohorts
Key enabling features:
• High throughput
• 12 Illumina indexes / up to 96 samples per run
• 16 SOLiD barcodes / up to 128 samples per run
• Only pay what you capture, scalable from 0.2 to 6.8Mb (sweet spot for 3rd Gen Seq)
• <0.2 Mb
• 0.2 – 0.5Mb
• 0.5 – 1.5Mb
• 1.5 – 3Mb
• 3 – 6Mb
• Very reproducible, excellent allelic balance for accurate heterozygote calls
• Custom and catalog content (kinome)
• Automation (library prep and capture)
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 24
• Inherited loss-of-function mutations in the tumor suppressor genes BRCA1, BRCA2, and
multiple other genes predispose to high risks of breast and/or ovarian cancer. Cancer-
associated inherited mutations in these genes are collectively quite common, but individually
rare or even private.
• To determine whether massively parallel, “next-generation” sequencing would enable
accurate, thorough, and cost-effective identification of inherited mutations for breast and
ovarian cancer, we developed a genomic assay to capture [with Agilent’s custom SureSelect],
sequence, and detect all mutations in 21 genes, including BRCA1 and BRCA2, with inherited
mutations that predispose to breast of ovarian cancer.
• There were zero false-positive calls of nonsense mutations, frameshift mutations, or
genomic rearrangements for any gene in any test sample.
• This approach enables widespread genetic testing and personalized risk assessment for
breast and ovarian cancer.
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 25
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 26
Excellent allelic balance
Deletion up to 19bp
eArray: a free tool to design custom SureSelect libraries
Customer A
Genomic locations #1Design parameters
Genomic locations #2Design parameters
Genomic locations #3Design parameters
Baits #1
Baits #2
Baits #3
Virtual bait library
Bait design
Bait design
Bait design
Librarydesign
Kit size
Quote
eArray Webportal
Customer B
Real DNA bait library
RNA bait library
Kit
Send to ManufacturingPass on Kit size info, etc.
Assemble kit
Ship Kit to Customer
Order library
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 27
Up to 55,000 Unique Baits/Kit
eArray – Supported species
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 28
H. sapiens, M. musculus, R. norvegicus, D. melanogaster, C. elegans, C. familiaris, S. cerviseae, S. pombe, G.
gallus, B. taurus, A. thaliana
eArrayXD – Import your own genomes
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 29
Available now
Indexing/Barcoding Procedure with SureSelect
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 30
Similar protocol available for SOLiD (x16 primers)
Illumina / SOLiD primer
SureSelect Pre-Capture
Primer
Prepared Library without Index Tag
2. Pre-Capture PCR
4. Post-Capture PCR
SureSelect Universal Primer
Illumina / SOLiD
Index Primer (Up to 12/16)
3. SureSelectHybridization/Capture
IndexCaptured
Library with Index Tag
1. Library prep • For optimum performance:
• Capture
• Index
• Pool
• Sequence
• Combine multiple samples per
sequencing lane
• Save on capture costs with
production scale
• Pay only for the Mb you capture:
• <0.2Mb 12-16 samples
• 0.2 – 0.5Mb
• 0.5 – 1.5Mb
• 1.5 – 3Mb 3-4 samples
• 3 – 6.8Mb
0.900.79
1.17
1.05
1.05
1.15
0.96
1.021.05
0.93
0.90
1.20
0.86
0.98
1.03
0.96
Standard Index Representation in Single SOLiD Quad
BC1
BC2
BC3
BC4
BC5
BC6
BC7
BC8
BC9
BC10
BC11Fold Representation Relative to the Mean
SOLiD Barcoding –16 barcodes of 0.2Mb Capture in 1 SOLiD Quad, 1x50bp, 30M reads
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 31
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Percentage reads in targeted regions:
Percentage reads in regions +/-100bp:
Percentage reads in regions +/-200bp:
HQ Reads
Mean/Barcode 1,844,819
Median/Barcode 1,843,950
Total/Quad 29,517,104
SOLiD Barcoding – Comparison with dbSNP16 barcodes of 0.2Mb Capture in 1 SOLiD Quad, 1x50bp, 50M reads
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 32
• Comparison of observed SNP calls vs. dbSNP 130
• We see 100% concordance rate with dbSNP across all called SNPs.
• There are, on average, ~80 novel SNPs called for each sample.
http://www.broadinstitute.org/gsa/wiki/index.php/The_Genome_Analysis_Toolkit
199 201 205 203 206 206 202 201 202 204 199 204 199 200 200 199
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
85 8797 97 91
108101 96 99
108
7999
96 10287 92
0
50
100
150
200
250
300
350
BC1 BC2 BC3 BC4 BC5 BC6 BC7 BC8 BC9 BC10 BC11 BC12 BC13 BC14 BC15 BC16
Nu
mb
er
of
SNP
sit
es
Barcode
dbSNP concordance across multiple barcodes
Novel Sites
dbSNP Mismatches
dbSNP Concordant
Agenda:
• Introduction
• Applications for Mendelian diseases research
• Custom biomarker discovery and profiling
• Applications for cancer biomarker discovery and profiling
• Automation
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 33
Cost: € 40,000 per patient/year
Targeted Therapies: Expensive, Can they be more effective?
100 %
0 %
Survival
Time (months)
Tito Fojo and Christine Grady, J Natl Cancer Inst. 2009;10:1044-8
Slide courtesy of Rene BernardsAgilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing Workflow
Herceptin
KRAS mutation alters response to EGFR therapy
in Colon Cancer
Mutant KRAS+EGFR-EGFR
Wild type KRAS+EGFR-EGFR
Amado et al. J Clin Oncol; 26:1626-1634 2008
KRAS mut: 32% PIK3CA mut: 13%
BRAF mut: 10%
Slide courtesy of Rene BernardsAgilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing Workflow
Current clinically useful correlations between kinome
alterations and drug responses
• HER2 expression (breast) Herceptin
• BCR-ABL translocation (CML) Gleevec
• KRAS mutation (colon) (no response) Cetuximab
• EGFR mutation (lung) Erlotinib
Somewhat further away, but promising:
• BRAF mutation (melanoma) PLX4032
• ALK mutation/translocation (lung, neuroblastoma) PF02341066
• PIK3CA mutation (breast) (no response) Herceptin
Slide courtesy of Rene BernardsAgilent SureSelect™ Platform
Enabling Products for the Next-Generation Sequencing Workflow
SureSelect “kinome” – Discovery and profiling of
biomarkers related to disease and/or drug response
• 518 putative kinases
• 12 PI3K domain-containing genes
• 6 PI3K regulatory components
• 13 diglyceride kinases
• 18 genes frequently mutated in human cancer
• 19 genes specifically known to be mutated in breast cancer
G. Manning et al Science 298 1912 (2002)
Slide courtesy of Rene BernardsAgilent SureSelect™ Platform
Enabling Products for the Next-Generation Sequencing Workflow
Kinome Kit Performance –3-5 samples per GAIIx lane / SOLID quad
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 38
Agenda:
• Introduction
• Applications for Mendelian diseases research
• Custom biomarker discovery and profiling
• Applications for cancer biomarker discovery and profiling
• Automation
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 39
Illumina Library Prep and SureSelect Enrichment on
the Bravo Automated Liquid Handling Platform
Page 40
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing Workflow
Next-Gen Sequencing and SureSelect Overview
Shear
Genomic DNA
Repair Ends
3’-dA Addition
Adapter
Ligation
PCR
Enrichment
Prepped
Library
SureSelect
Oligo Capture
LibraryLibrary
Hybridization
Magnetic
Bead Capture
PCR
QA
Illumina
Cluster &
Sequence
Library Prep
SureSelect
Target Enrichment
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 41
Next-Gen Sequencing and SureSelect Overview
Shear
Genomic DNA
Repair Ends
3’-dA Addition
Adapter
Ligation
PCR
Enrichment
Prepped
Library
SureSelect
Oligo Capture
LibraryLibrary
Hybridization
Magnetic
Bead Capture
PCR
QA
Illumina
Cluster &
Sequence
Library Prep
SureSelect
Target Enrichment
Agilent Automation
Bravo Liquid Handler
Automated
Bead Capture
Automated
Purification
Automated
Purification
Automated
Purification
Automated
Purification
Automated
Purification
Automated
Purification
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 42
Most Comprehensive, Ever Expanding SureSelect™ Menu
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 43
ProductTarget amount (catalog
number)Reactions/kit Product Definition
X-demo 3.4 Mb 5 Exons in the human X-chr
All Exon v1 38 Mb 5-10,000Catalog content from CCDS + >1000
ncRNA
All Exon Plus38 Mb + up to 6.8 Mb of
custom content5-10,000
Add custom content to All Exoncatalog content
All Exon v2 38 Mb + RefSeq 5-10,000CCDS Sept. 2009
+ additional RefSeq
All Exon 50Mb 50Mb 5-10,000GENCODE content – Most comprehensive coverage
Multiplexable
Kinome <3Mb 5-10,000 All kinases
Indexed custom content
<0.2 Mb, 0.2-0.49 Mb, 0.5-1.49 Mb, 1.5- 2.9 Mb 3 – 6.8 Mb
10 – 5,000Cost-saving custom offering –
Illumina (12 indexes) and SOLiD (16 barcodes)
Summary: Efficient Enrichment for Re-Sequencing
• Most comprehensive Human All Exon catalog products
• New 50Mb catalog content
• Multiplexable (HiSeq and SOLiD4)
• With option to add up to 6Mb custom (Exon Plus)
• Enables biomarker discovery and profiling
• New Kinome catalog content and custom content
• Indexing/Barcoding for Illumina and SOLiD
• Scalable and affordable from 0.2 – 6.8Mb
• Free web portal, eArray, enables fully custom design
• Fastest way to biological answer
• Low DNA input
• Accurate SNP calls
• Fast, reproducible and, automatable
• Available for SE and PE on Illumina and SOLiD
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 44
Agilent SureSelect™ Platform
Enabling Products for the Next-Generation
Sequencing WorkflowPage 45
Acknowledgements
• Collaborators:
o Broad Institute
• Chad Nussbaum et. al
• Stacey Gabriel et. al
• Sheila Fisher et. al
o Sanger Institute
• Daniel Turner et.al.,
o NKI
• Rene Bernards
• Ian Majewski
o All our early access collaborators (over 20
institutions worldwide)
•Life Technologies and Illumina R&D/Marketing