title: Whole-Genome Sequencing of a Single Viral Species from a Highly Heterogeneous Sample.
abstract: Metagenomic studies suggest that only a small fraction of the viruses that exist in nature have been identified and studied. Characterization of unknown viral genomes is hindered by the many genomes populating any virus sample. A new method is reported that integrates drop-based microfluidics and computational analysis to enable the purification of any single viral species from a complex mixed virus sample and the retrieval of complete genome sequences. By using this platform, the genome sequence of a 5243 bp dsDNA virus that was spiked into wastewater was retrieved with greater than 96% sequence coverage and more than 99.8% sequence identity. This method holds great potential for virus discovery since it allows enrichment and sequencing of previously undescribed viruses as well as known viruses.
authors: Department of Physics, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138 (USA).
authors: Hee-Sun
authors: HS
authors: Han
authors: Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260 (USA).
authors: Paul G
authors: PG
authors: Cantalupo
authors: Department of Physics, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138 (USA).
authors: Assaf
authors: A
authors: Rotem
authors: Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260 (USA).
authors: Shelley K
authors: SK
authors: Cockrell
authors: Department of Physics, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138 (USA).
authors: Martial
authors: M
authors: Carbonnaux
authors: Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260 (USA).
authors: James M
authors: JM
authors: Pipas
authors: Department of Physics, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138 (USA). weitz@seas.harvard.edu.
authors: David A
authors: DA
authors: Weitz
mesh_terms: D001483:Base Sequence; D004267:DNA Viruses / Q000235:genetics* / Q000302:isolation & purification*; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D046210:Microfluidic Analytical Techniques / Q000379:methods*
chemical_list: D004279:DNA, Viral
keywords: genome sequencing; high-throughput screening; microemulsions; microfluidics; viruses