BME 130 – Genomes Lecture 4 Sequencing technology II Next generation sequencing.

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Presentation transcript:

BME 130 – Genomes Lecture 4 Sequencing technology II Next generation sequencing

Figure 4.2 Genomes 3 (© Garland Science 2007)

Figure 4.3a Genomes 3 (© Garland Science 2007)

Roche / 454

454 Process Overview 1) Prepare Adapter Ligated ssDNA Library 2) Clonal Amplification on 28 µ beads 4) Perform Sequencing by synthesis on the 454 Instrument of ~250,000 molecules 3) Load beads and enzymes in PicoTiter Plate™

Key sequence = TCAG for identifying wells and calibration TACGTACG Flow Order 454 base calling

Limitations of Roche/454

Solexa / Illumina

Limitations of Solexa/Illumina

ABI/SOLiD

Limitations of ABI/SOLiD

Applications of high-throughput sequencing

Genome assembly

CHiP-Seq Johnson et al., Science 8 June 2007: Vol no. 5830, pp

RNASeq Cloonan et al., Genome Biology 2008, 9:234

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