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Efficient de novo assembly of single-cell bacterial genomes from short-read data sets.

Whole genome amplification by the multiple displacement amplification (MDA) method allows sequencing of DNA from single cells of bacteria that cannot be cultured. Assembling a genome is challenging, however, because MDA generates highly nonuniform coverage of the genome. Here we describe an algorithm tailored for short-read data from single cells that improves assembly through the use of a progressively increasing coverage cutoff. Assembly of reads from single Escherichia coli and Staphylococcus aureus cells captures >91% of genes within contigs, approaching the 95% captured from an assembly based on many E. coli cells. We apply this method to assemble a genome from a single cell of an uncultivated SAR324 clade of Deltaproteobacteria, a cosmopolitan bacterial lineage in the global ocean. Metabolic reconstruction suggests that SAR324 is aerobic, motile and chemotaxic. Our approach enables acquisition of genome assemblies for individual uncultivated bacteria using only short reads, providing cell-specific genetic information absent from metagenomic studies.

Pubmed ID: 21926975


  • Chitsaz H
  • Yee-Greenbaum JL
  • Tesler G
  • Lombardo MJ
  • Dupont CL
  • Badger JH
  • Novotny M
  • Rusch DB
  • Fraser LJ
  • Gormley NA
  • Schulz-Trieglaff O
  • Smith GP
  • Evers DJ
  • Pevzner PA
  • Lasken RS


Nature biotechnology

Publication Data

October 14, 2011

Associated Grants

  • Agency: NHGRI NIH HHS, Id: 2 R01 HG003647
  • Agency: NCRR NIH HHS, Id: 3P41RR024851-02S1
  • Agency: NCRR NIH HHS, Id: P41 RR024851
  • Agency: NHGRI NIH HHS, Id: R01 HG003647

Mesh Terms

  • Algorithms
  • Bacteria
  • Base Sequence
  • Contig Mapping
  • Databases, Nucleic Acid
  • Deltaproteobacteria
  • Escherichia coli
  • Genome, Bacterial
  • Likelihood Functions
  • Sequence Analysis, DNA
  • Single-Cell Analysis
  • Staphylococcus aureus