Genome sequencing in microfabricated high-density picolitre reactors View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2005-09

AUTHORS

Marcel Margulies, Michael Egholm, William E. Altman, Said Attiya, Joel S. Bader, Lisa A. Bemben, Jan Berka, Michael S. Braverman, Yi-Ju Chen, Zhoutao Chen, Scott B. Dewell, Lei Du, Joseph M. Fierro, Xavier V. Gomes, Brian C. Godwin, Wen He, Scott Helgesen, Chun He Ho, Gerard P. Irzyk, Szilveszter C. Jando, Maria L. I. Alenquer, Thomas P. Jarvie, Kshama B. Jirage, Jong-Bum Kim, James R. Knight, Janna R. Lanza, John H. Leamon, Steven M. Lefkowitz, Ming Lei, Jing Li, Kenton L. Lohman, Hong Lu, Vinod B. Makhijani, Keith E. McDade, Michael P. McKenna, Eugene W. Myers, Elizabeth Nickerson, John R. Nobile, Ramona Plant, Bernard P. Puc, Michael T. Ronan, George T. Roth, Gary J. Sarkis, Jan Fredrik Simons, John W. Simpson, Maithreyan Srinivasan, Karrie R. Tartaro, Alexander Tomasz, Kari A. Vogt, Greg A. Volkmer, Shally H. Wang, Yong Wang, Michael P. Weiner, Pengguang Yu, Richard F. Begley, Jonathan M. Rothberg

ABSTRACT

The proliferation of large-scale DNA-sequencing projects in recent years has driven a search for alternative methods to reduce time and cost. Here we describe a scalable, highly parallel sequencing system with raw throughput significantly greater than that of state-of-the-art capillary electrophoresis instruments. The apparatus uses a novel fibre-optic slide of individual wells and is able to sequence 25 million bases, at 99% or better accuracy, in one four-hour run. To achieve an approximately 100-fold increase in throughput over current Sanger sequencing technology, we have developed an emulsion method for DNA amplification and an instrument for sequencing by synthesis using a pyrosequencing protocol optimized for solid support and picolitre-scale volumes. Here we show the utility, throughput, accuracy and robustness of this system by shotgun sequencing and de novo assembly of the Mycoplasma genitalium genome with 96% coverage at 99.96% accuracy in one run of the machine. More... »

PAGES

376

References to SciGraph publications

Journal

TITLE

Nature

ISSUE

7057

VOLUME

437

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  • Electron Beam Sculpting Of Tunneling Junction For Nanopore Dna Sequencing
  • Methods Of Selecting Biologic-Producing Cell Lines By Next Generation Sequencing
  • Digital Analyte Analysis
  • Data Processing System And Methods
  • Novel Macrocyles And Methods For Their Production
  • Method For Identifying The Sequence Of One Or More Variant Nucleotides In A Nucleic Acid Molecule
  • Methods For Accurate Sequence Data And Modified Base Position Determination
  • Paired End Bead Amplification And High Throughput Sequencing
  • Method And System For Delta Double Sampling
  • Integrated Sample Preparation Systems And Stabilized Enzyme Mixtures
  • Methods For Estimating Genome-Wide Copy Number Variations
  • Detecting And Classifying Copy Number Variation
  • High Throughput Genome Sequencing On Dna Arrays
  • Chemical Sensors With Consistent Sensor Surface Areas
  • Methods To Diagnose And Immunize Against The Virus Causing Human Merkel Cell Carcinoma
  • Biological Analysis Arrangement And Approach Therefor
  • System And Method For Producing Functionally Distinct Nucleic Acid Library Ends Through Use Of Deoxyinosine
  • Methods And Apparatus For Measuring Analytes
  • Detection And Quantification Of Biomolecules Using Mass Spectrometry
  • Chemical Sensor With Sidewall Spacer Sensor Surface
  • Method For Determining Fetal Chromosomal Abnormality
  • Methods And Solutions For Inhibiting Undesired Cleaving Of Labels
  • Massive Parallel Method For Decoding Dna And Rna
  • Oligomer Sequences Mapping
  • Four-Color Dna Sequencing By Synthesis Using Cleavable Fluorescent Nucleotide Reversible Terminators
  • Methods And Apparatus For Measuring Analytes Using Large Scale Fet Arrays
  • Composition, Method And Kit For Detecting Bacteria By Means Of Sequencing
  • Chemical Sensor With Protruded Sensor Surface
  • Solid Support For High-Throughput Nucleic Acid Analysis
  • Nanogap Transducers With Selective Surface Immobilization Sites
  • Methods For Forming Mixed Droplets
  • Piscine Reovirus Diagnostic Compositions
  • Nucleic Acid Analysis Using Sequence Tokens
  • Method For Identifying The Sequence Of One Or More Variant Nucleotides In A Nucleic Acid Molecule
  • Methods And Apparatus For Measuring Analytes Using Large Scale Fet Arrays
  • Models For Analyzing Data From Sequencing-By-Synthesis Operations
  • Programmable Oligonucleotide Synthesis
  • Massive Parallel Method For Decoding Dna And Rna
  • Assay Methods Using Nicking Endonucleases
  • One-Transistor Pixel Array
  • Electron Beam Sculpting Of Tunneling Junction For Nanopore Dna Sequencing
  • Active Chemically-Sensitive Sensors With Reset Switch
  • Manufacturable Sub-3 Nanometer Palladium Gap Devices For Fixed Electrode Tunneling Recognition
  • Massively Parallel Single Cell Analysis
  • Method For Sequencing A Polynucleotide Template
  • Methods, Compounds And Systems For Detecting A Microorganism In A Sample
  • Method Of Performing One-Step, Single Cell Rt-Pcr
  • Dna Sequencing Using Multiple Metal Layer Structure With Different Organic Coatings Forming Different Transient Bondings To Dna
  • Amplification Primers And Methods
  • Massive Parallel Method For Decoding Dna And Rna
  • Method For Generating Clonotype Profiles Using Sequence Tags
  • Variant Database
  • Nucleic Acid Interaction Analysis
  • Massively Parallel Single Cell Analysis
  • Analysis Methods
  • Efficient Arrays Of Amplified Polynucleotides
  • Second Harmonic Imaging Nanoprobes And Techniques For Use Thereof
  • Method And System For Delta Double Sampling
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nature03959

    DOI

    http://dx.doi.org/10.1038/nature03959

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1021574562

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/16056220


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