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DNA sequencing: a generation game Dr John Milton VP Research, Oxford Nanopore Technologies

DNA sequencing: a generation game Dr John Milton VP Research, Oxford Nanopore Technologies. Today. The DNA sequencing explosion How applications have evolved How users have evolved Today’s technologies Tomorrow’s technologies. From discovery to technology explosion. $ human Genome

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DNA sequencing: a generation game Dr John Milton VP Research, Oxford Nanopore Technologies

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  1. DNA sequencing: a generation gameDr John MiltonVP Research, Oxford Nanopore Technologies

  2. Today • The DNA sequencing explosion • How applications have evolved • How users have evolved • Today’s technologies • Tomorrow’s technologies CONFIDENTIAL

  3. From discovery to technology explosion $ human Genome $3 billion $2-3 million $250k $50k $20k ?<$5k? • 1868: Discovery of DNA • 1953: Watson and Crick propose double helix structure • 1977: Sanger sequencing • 1985: PCR • 2000: Working draft human genome announced (Sanger method) • 2005: 454 sequencer launch (pyrosequencing) • 2006: Genome Analyzer launched (Solexa sequencing) • 2007: SOLiD launched (ligation sequencing) • 2009: Whole human genome no longer merits Nature/Science paper • 2010: “third-gen” systems on the horizon

  4. Volume of DNA data: the early years

  5. Sequence data outputUK’s Sanger Centre, 2007-today March 2010 Sept 2007 Sept 2008 Sept 2009 CONFIDENTIAL

  6. © Macmillan: Nature 1st April 2010 CONFIDENTIAL

  7. ‘next gen’ systems today (UK) Source: http://pathogenomics.bham.ac.uk/hts/

  8. ‘next gen’ systems today (US) CONFIDENTIAL

  9. Turning DNA data into knowledge Wheat Tomato Chimpanzee Human Microbiome Project Neanderthal Chicken Corn Anthrax Bovine Malaria parasite Panda Arabidopsis

  10. Which applications are labs performing? Source: GenomeWeb Survey 2010 CONFIDENTIAL

  11. The rise of epigenetics Genome “what may happen” Epigenetics “how ‘what may happen’ may vary” 3rd dimension of the genome Gene expression / protein analysis “what actually happened” In this cell, at this time CONFIDENTIAL

  12. Technologies CONFIDENTIAL

  13. Implications of advancing technology Expansion in computing required to capture all variation and interpret its meaning  democratisation of DNA information CONFIDENTIAL

  14. DNA sequencing generations Then + Now Now Now + anticipated Anticipated Next -single mol AND electronic 1st Gen Sanger 2nd Gen -parallised 2nd Gen -single mol or electronic • Low • throughput • High cost • Accurate • Broad user • base • Optical • Amplification needed • Highly parallel • Improved cost and • Throughput • More centralised • users • Optical • Single-molecule • Highly parallel • Cost similar • New applications • Or electronic, • clonal • Direct electrical (no optics) • Single-molecule, highly parallel • Transformation of workflow • Designed to broaden user base, • deliver step change in cost, power • New applications Sanger GAII (Solexa/Illumina) SOLiD (Agencourt/LIFE) FLX (454/Roche) Helicos Pacific Biosciences Ion Torrent (LIFE Starlight) Nanopores Estimated cost of a human genome using these technologies $70M $200k --- $50k ---- $20k --- 15k--- ?$5k - $?

  15. Sanger Sequencing (CE) 3730: “workhorse of the HGP” Human genome $ 100millions CONFIDENTIAL

  16. Illumina GA (‘Solexa sequencing’)sequencing by synthesis Human genome $ 50k+ Hi-Seq falling through $20k

  17. SOLiD: sequencing by ligation Human genome $ 50k+ CONFIDENTIAL

  18. 454: pyrosequencing Human genome $ ‘000ks Long read lengths: strong for clinical CONFIDENTIAL

  19. Helicos: Heliscope Single molecule genome: $ 50k reagents Machine $750k CONFIDENTIAL

  20. Coming soon Ion Torrent $50k Clonal Short run time Errors? Yield? Pacific Biosciences $750k Single molecule Long read lengths Errors? Yield? Single molecule Electronic CONFIDENTIAL

  21. What is a nanopore? • Nanopore = ‘very small hole’ • Electrical current flows through the hole • Introduce analyte of interest into the hole  identify “analyte” by the disruption or block to the electrical current Current flow Electronic & Single molecule out current in 21 CONFIDENTIAL

  22. Engineering nature’s nanopores CONFIDENTIAL

  23. Nucleotide Recognition 23

  24. Movie can be found at: http://www.nanoporetech.com/sequences 24 CONFIDENTIAL

  25. Platform Technology DNA Sequencing Proteins Polymers SmallMolecules Adaptable protein nanopore: Application Specific Sensor array chip: many nanopores in parallel Generic Platform Electronic read-out system 25

  26. Ultimately: will we sequence every person? Every cancer: Accurate diagnosis and targeted treatment? Every baby: Lifetime ‘baseline’ resource, disease prevention? Every infectious agent: Control of disease spread and resistance

  27. Ultimately: will we sequence every species? http://seedmagazine.com/interactive/genome/ 1995 2005 2002 2000 2002 2009

  28. Thank youquestions?

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