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Second and third generation sequencing technologies

Second and third generation sequencing technologies. Morten Rasmussen, PhD. Outline. What was before? All the new players 454 Illumina SOLiD Helicos PGM Pacific Biosciences Oxford Nanopores Many more to come. What is first generation sequencing?. Sequencing = read DNA code, A-C-G-T

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Second and third generation sequencing technologies

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  1. Centre for GeoGenetics Second and third generation sequencing technologies Morten Rasmussen, PhD

  2. Centre for GeoGenetics Outline • What was before? • All the new players • 454 • Illumina • SOLiD • Helicos • PGM • Pacific Biosciences • Oxford Nanopores • Many more to come

  3. Centre for GeoGenetics What is first generation sequencing? • Sequencing = read DNA code, A-C-G-T • Sanger sequencing, 1977 • Dideoxy chemistry • Read length 700-1000bp • Requires PCR primer sites (>40bp) • Often requires cloning

  4. Centre for GeoGenetics What is first generation sequencing?

  5. Centre for GeoGenetics Introduction – second generation sequencingWhat is second generation sequencing? 1977 2006 x very big number x 1 2007 1 - 384 105 - >109 x even bigger number 1998 2008 x gigantic number x 384 2011 x big number

  6. Centre for GeoGenetics Introduction – second generation sequencingDefining features of second generation sequencing • Library • Clonal Amplification • Massively Parallel Sequencing DNA from extract Polished DNA Library Adaptors Library molecule

  7. Centre for GeoGenetics Second generation sequencing platforms 454 Illumina SOLiD Ion Torrent (PGM)

  8. Centre for GeoGenetics The GS FLX (454) platform • Clonal Amplification • Massively parallel sequencing

  9. Centre for GeoGenetics The GS FLX (454) platform

  10. Centre for GeoGenetics Ion Torrent and Proton

  11. Centre for GeoGenetics Introduction – second generation sequencingThe Illumina (GAIIx) platform • Clonal Amplification • Massively parallel sequencing

  12. Centre for GeoGenetics Illumina reaction flow P5 Index P7 Insert Read1 Read2 ReadIndex

  13. Centre for GeoGenetics Sequencing by Oligo Ligation and Detection

  14. Centre for GeoGenetics SOLiD 2-base encoding

  15. Centre for GeoGenetics Third generation sequencing • Defined by no amplification • Simple sample preparation Oxford Nanopore Helicos PacBio

  16. Centre for GeoGenetics Helicos

  17. Centre for GeoGenetics Pacific Biosciences - SMaRT

  18. Centre for GeoGenetics Oxford Nanopore Video

  19. Centre for GeoGenetics Oxford Nanopore

  20. Centre for GeoGenetics Current status (Mar 2012)

  21. Centre for GeoGenetics Current status part 2

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