Genome sequencing in microfabricated high-density picolitre reactors. Academic Article uri icon

Overview

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.

authors

publication date

  • July 31, 2005

Research

keywords

  • Genome, Bacterial
  • Genomics
  • Microchemistry
  • Mycoplasma genitalium
  • Sequence Analysis, DNA

Identity

PubMed Central ID

  • PMC1464427

Scopus Document Identifier

  • 24044455869

Digital Object Identifier (DOI)

  • 10.1038/nature03959

PubMed ID

  • 16056220

Additional Document Info

volume

  • 437

issue

  • 7057