Exploring Illumina’s Newest Sequencing Solutions
Exploring Illumina’sNewest SequencingSolutionsAppliedGenomicsDan GhebaSr. Sequencing SpecialistSeptember 14th, 2016dgheba@illumina.com 2016 Illumina, Inc. All rights reserved.Illumina, 24sure, BaseSpace, BeadArray, BlueFish, BlueFuse, BlueGnome, cBot, CSPro, CytoChip, DesignStudio, Epicentre, ForenSeq, Genetic Energy, GenomeStudio, GoldenGate, HiScan, HiSeq,HiSeq X, Infinium, iScan, iSelect, MiniSeq, MiSeq, MiSeqDx, MiSeq FGx, NeoPrep, NextBio, Nextera, NextSeq, Powered by Illumina, SureMDA, TruGenome, TruSeq, TruSight, Understand YourGenome, UYG, VeraCode, verifi, VeriSeq, the pumpkin orange color, and the streaming bases design are trademarks of Illumina, Inc. and/or its affiliate(s) in the US and/or other countries. All othernames, logos, and other trademarks are the property of their respective owners.1
HiSeq HD Systems2
2014: HiSeq XPopulation Scale Human Genome Sequencing1.8T 6B READS PE150 3 DAYS3
Sequencing Power for Every ScaleIncreasing system output & priceWGS – Large ScaleHiSeq X TenRNA-seqExomesWGSMethylation1800 Gb 6B2x150HiSeq X FiveHiSeq 40001500 Gb 5B2x150ProkaryoteTargetedQCHiSeq 3000HiSeq 2500NextSeqMiSeqMiniSeq120 Gb 400M2x1501000 Gb 4B2x125750 Gb 2.5B2x15015 Gb 25M2x3007.5 Gb 25M2x150Decreasing price per GB41800 Gb 6B2x150
HiSeq 4000:Over 2x more data in 1/3 the time!1.5 TB 5B READS PE1503.5 DAYS600 GB 3B READS PE10011 DAYSV351 TB 4B READS PE1256 DAYSV4HD
What can you do with just ONE LANE of HiSeq 4000 data?6MouseArabidopsisDrosophila2.7 Gb Genome157 Mb Genome130 Mb Genome34X Coverage590X Coverage715X Coverage
Innovative Patterned Flow Cell TechnologyPatterned FCNon-patterned FCDefined featureOrdered spacingUndefined featureRandom spacingNanowell SubstrateBillions of Ordered WellsDefined feature sizeOptimal cluster spacingIncreased cluster densityFaster, Simplifiedimaging7
Supported Library Prep KitsOn HiSeq 3000 and 4000 SystemsDNATargeted DNARNA / RegulationTruSeq NanoNextera Rapid Capture ExomeTruSeq Stranded mRNATruSeq PCR-freeNextera Rapid Capture CustomTruSeq Stranded Total RNANextera XTTruSeq ExomeTruSeq RNA AccessNextera Mate-PairTruSeq Rapid ExomeTruSeq Small RNATruSeq Synthetic Long ReadTruSeq ChIPTruSeq DNA Methylation8
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ATAC-seq on the Hiseq 4000Data from Eric Chow, Ph.D., UCSF10
ATAC-seq on the Hiseq 4000Data from Eric Chow, Ph.D., UCSF11
Robust performance of patterned flow cells overa broad range of input concentrations withquality librariesSpecification is 312 M reads per lane75% PF yields 368 M reads per lane12
Nextseq 50013
The NextSeq 500 Delivers on Three Key Aspects Flexibility Speed Simplicity14
System enhancements: OpticsPower of high throughput appsSize and affordability of a desktop sequencer.6xMiSeq imagingcapability6 parallel miniaturized,solid-state optics modules151/3Size of a HiSeq1/3Capital costof a HiSeq
Two Channel SBS – NextSeq 500Two channel SBS uses 2 imagesBuilds template over 5 cyclesClusters appearing in green only are TClusters appearing in red only are CClusters appearing in both images are AClusters not present in either green norred are GCluster intensities are plotted and basesare called accordingly16
Fast 2-channel SBS25%24 hfaster18 hMiSeq v3NextSeq 5002x75bp runFASTER CHEMISTRYHALF THE IMAGES17
18Human Genome30 HOURSExome Transcriptome18 HOURSGene Expression Profile12 HOURS2 x 150bp2 x 75bp1 x 75bp
One System, Two Output ModesHighMid120 GIGABASES PE150400M CLUSTERS40 GIGABASES PE150130M CLUSTERS.19
Flexible applicationsHigh-OutputUp to 120 GbUp to 40 Gb400M clusters PF1 x 75 bp, 2x75 bp, 2 x 150 bp130M clusters PF2 x 75 bp to 2 x 150 -Seq20GEXprofiles20Mid-Output6-36pan
Sep 02, 2016 · TruSeq Stranded mRNA TruSeq Stranded Total RNA TruSeq RNA Access TruSeq Small RNA TruSeq ChIP TruSeq DNA Methylation DNA Targeted DNA RNA / Regulation Supported Library Prep Kits On HiSeq 3000 and 4000 Systems. 9. 10 ATAC-s
sequencing. By modifying the primer constructs to include the Illumina adapters and a unique barcode, the amplified region of the 16S rRNA gene can be identified in a pooled sample and the sample is prepared for sequencing with the Illumina MiSeq platform. Figure 2. Protocol for barcoded Illumina sequencing. A target gene is identified, which .
(Next-Generation Sequencing) 9/12/16 10 Impact of Next-Generation Sequencing Illumina NextSeq Flow Cell 400,000,000 DNA fragments X 300 nucleotides 120,000,000,000 nucleotides in 29 hours . 9/12/16 11 Illumina Sequencing Benchmarks NextSeq 500 1 Technician 29 hours 4000
Also referred to as Next-Generation Sequencing Parallelize the sequencing process, producing thousands or millions of sequences concurrently Lower the cost of DNA sequencing beyond what is possible with standard dye-terminator methods. In ultra-high-throughput sequencing as many as 500,000 sequencing-by-synthesis operations may
The mRNA library was prepared using an Illumina Truseq Stranded mRNA High Throughput Prep kit and samples were sequenced using an Illumina NextSeq 500 Mid-Output Sequencing Reagent kit (v2, 150 cycles), 132 M reads on an Illumina NextSeq 5
From library prep, arrays, and sequencing to informatics, Illumina genomic solutions empower researchers and clinical researchers across the globe to find the answers they seek. When you join the Illumina community, you become part of a dynamic scientific movement that includes thousands of researchers and industry thought leaders.
Next generation sequencing (NGS) Nature Methods 2011 Sequencing technology defies Moore's law. 2009: Illumina, Helicos 40-50K 4 2010: 5K , a few days Sequencing the Human Genome Year Log 10 (price) 2000 2005 2010 10 8 6 60K , 2 weeks 4 2 2014: 1000 , 24 hrs 2008: ABI SOLiD 2007: 454
2 Part # 15045845_Rev.D FOR RESEARCH USE ONLY By the end of this training, you will be able to: –List the major steps in the Illumina sequencing workflow –Describe cluste
the Tiny seed Sequencing Use sensory trays/sequencing cards to explore the story the Tiny seed Sequencing Use sensory trays/sequencing cards to explore the story . Using sequencing cards to plant sunflower seeds Make butterfly Plant peas and carrots as part of a food farm Create habitats using a variety of
