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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.
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:
There are four technologies that were talked about today.