GENOME SEQUENCING. I. Genome sequencing The Sanger Method (1977) Denaturation +priming Polymerization

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  • GENOME SEQUENCING
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  • I. Genome sequencing The Sanger Method (1977) Denaturation +priming Polymerization
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  • End of polymerization I. Genome sequencing The Sanger Method (1977)
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  • I. Genome sequencing The Sanger Method (1977)
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  • I. Genome sequencing The caveats of the Sanger Method Technically cumbersome and time consuming (limited samples processing) High amounts of starting material Not well-suited for high-throughput analyses (genome sequencing)
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  • 1st generation sequencing : Improvement of sequence analysis (column electrophoresis) I. Genome sequencing The Sanger Method (1977)
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  • 1st generation sequencing : Improvement of sequence analysis (capillary electrophoresis) I. Genome sequencing The Sanger Method (1977)
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  • Parallel random sequencing : after fragmentation and cloning 1st generation sequencing : Improvement of the methodology I. Genome sequencing
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  • 2nd generation sequencing : Areas of improvement I. Genome sequencing
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  • 2nd generation sequencing : Improvement of template preparation : amplification by PCRPCR Emulsion PCR (emPCR) I. Genome sequencing Used by 454 (life science), Polonator, Ion torrent and Solid
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  • 2nd generation sequencing : Improvement of template preparation : amplification by PCRPCR Bridge PCR (illumina plateform)illumina plateform I. Genome sequencing
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  • 2nd generation sequencing : Improvement of template preparation : Single Molecule Sequencing I. Genome sequencing
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  • Sequencing by reversible terminator (illumina/Solexa genome Analyzer) 2nd generation sequencing : real-time sequencing I. Genome sequencing
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  • Sequencing by reversible terminator Used in combination with bridge PCR in the illumina/Solexa genome Analyzer 2nd generation sequencing : real-time sequencing I. Genome sequencing
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  • Pyrosequencing 2nd generation sequencing : real-time sequencing I. Genome sequencing Used in the 454 plateform (Life Sciences) emPCR+Pyrosequencing
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  • Ligase-based sequencing 2nd generation sequencing : real-time sequencing I. Genome sequencing
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  • Ligase-based sequencing 2nd generation sequencing : real-time sequencing I. Genome sequencing SOLiD DNA Sequencing Technology (Applied Biosystems)
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  • SMRT Sequencing Technology : Single-Molecule Real-Time Sequencing A true continous imaging of dye-labelled nucleotides incorporation 2nd generation sequencing : real-time sequencing I. Genome sequencing Pacific BiosciencesPacific Biosciences : SMRT : Single Molecule Real-Time Sequencing
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  • I. Genome sequencing Today's technologies generate huge amount of short, high quality sequencing data
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  • II. Data analysis
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  • III. NGS Applications
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