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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.

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lecture_notes/mar31.1270084821.txt.gz · Last modified: 2010/03/31 18:20 by svasili