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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 == Pyrosequencing == == solid == == Illumina/solexa == == Ion Torrent == === Ideas for data storage layout in Campusrocks ===
