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Next Generation Sequencing
Ion Torrent Method
Overview
• Similar idea as SMRT
• Detects the
chemiluminescence given
off when a proton (H+) is
released
• Use ion chip
Library Construction
• If RNA, converted into cDNA
• DNA fragmentation
• Adapter ligated to DNA
• Denaturation
Image provi...
Template Prep/ePCR Amplification
• Fragments are attached to beads
• ePCR
Sequencing
• Based on standard pyrosequencing
• Measures light from chemiliuminescent
reagents by measuring direct release...
Data Analysis
• Output  FASTQ
Image provided by http://mendel.iontorrent.com/ion-docs/On-Dataflow.html
Work Cited
• Ambry Genetics. Making Sense of NextGent Sequencing. Kelly Gonzalez, MS,
CGC, and Senior Manager of Clinical ...
Recommended Video
See what you've been missing:
Explore RNA-Seq for Gene
Expression research
https://www.youtube.com/user/...
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Ion Torrent Sequencing

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This presentation covers how Ion Torrent Sequencing, a type of Next Generational Sequencing, works.

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Ion Torrent Sequencing

  1. 1. Next Generation Sequencing Ion Torrent Method
  2. 2. Overview • Similar idea as SMRT • Detects the chemiluminescence given off when a proton (H+) is released • Use ion chip
  3. 3. Library Construction • If RNA, converted into cDNA • DNA fragmentation • Adapter ligated to DNA • Denaturation Image provided by http://www.thermoscientificbio.com/ngs-library-preparation-kits/
  4. 4. Template Prep/ePCR Amplification • Fragments are attached to beads • ePCR
  5. 5. Sequencing • Based on standard pyrosequencing • Measures light from chemiliuminescent reagents by measuring direct release of H+ (protons) from the sequencing reaction
  6. 6. Data Analysis • Output  FASTQ Image provided by http://mendel.iontorrent.com/ion-docs/On-Dataflow.html
  7. 7. Work Cited • Ambry Genetics. Making Sense of NextGent Sequencing. Kelly Gonzalez, MS, CGC, and Senior Manager of Clinical Genomics. Available at http://www.ambrygen.com/sites/default/files/pdfs/NERRG_4-10- 12_Making_Sense_of_NetGen_Sequencing_KG(3).pdf. Access verified May 21, 2014. • Omixon. Allen Van Deynze, 2010; Solanaceae Coordinated Agricultural Project. Next Generation Sequencing. Available at http://www.omixon.com/the-basics- of-next-generation-sequencing/. Access verified May 21, 2014. • AllSeq. Life Technologies – Ion Torrent. http://allseq.com/knowledgebank/sequencing-platforms/life-technologies-ion- torrent Access verified Aug. 1, 2014.
  8. 8. Recommended Video See what you've been missing: Explore RNA-Seq for Gene Expression research https://www.youtube.com/user/ionto rrent

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