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Chemistry of Sequencing Technology

Nader talked about the chemistry behind 4 Sequencing Platforms : SOLiD Bioanalyzer, 454, Illumina/Solexa, and charge based detection system Now sequencing.

Sequencing workflow : Genomic DNA → fragment → amplification → immobilization → Sequencing.
Time required for sequencing through these platforms is :
1-2 hrs for CBD.
9 hrs for 454.
3-7 days for Illumina/Solexa.
7-10 days for SOLiD.

These platforms are classified into two :
1) Sequencing by synthesis (SBS) : 454, Illumina/Solexa, Helicos, Pacbio, Charge based detection system Now Sequencing.
2) Sequencing by hybridization : SOLiD.

Sequencing by synthesis
Involves synthesizing complementary strand of DNA to the template DNA strand to be sequenced. Four enzymes are used in this method - DNA polymerase, ATP sulfyrylase, Luciferase, and Apyrase. The light generated is captured as a signal which detects the base incorporated. The read length is approximately 400 nucleotides.

Alternative Notes

High Level Overview

The high level sequencing workflow for all next gen tools is as follows: Fragment Sample → Amplify Fragments → Sequence Fragments

There are two main flavors of next generation sequencing technology:

  • Sequencing by Synthesis (SBS)
  • Hybridization

Examination of Individual Technologies

There are four technologies that were talked about today.

  • Pyrosequencing (454)
  • solid
  • Illumina/solexa
  • Ion Torrent
Ion Torrent

Ideas for data storage layout in Campusrocks

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lecture_notes/mar31.1270090593.txt.gz · Last modified: 2010/03/31 19:56 by jstjohn