Sequencing of Genomes from Environmental Single Cells. In: Next Generation Sequencing. Methods in Molecular Biology, Vol. 1712

Bowers, Robert M.; Lee, Janey; Woyke, Tanja

doi:10.1007/978-1-4939-7514-3_8

Sequencing of Genomes from Environmental Single Cells. In: Next Generation Sequencing. Methods in Molecular Biology, Vol. 1712

Book ·

DOI:https://doi.org/10.1007/978-1-4939-7514-3_8· OSTI ID:1546616

Bowers, Robert M. ^[1]; Lee, Janey ^[1]; Woyke, Tanja ^[1]

USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)

Sequencing of single bacterial and archaeal cells is an important methodology that provides access to the genetic makeup of uncultivated microorganisms. We here describe the high-throughput fluorescence-activated cell sorting-based isolation of single cells from the environment, their lysis and strand displacement-mediated whole genome amplification. We further outline 16S rRNA gene sequence-based screening of single-cell amplification products, their preparation for Illumina sequencing libraries, and finally propose computational methods for read and contig level quality control of the resulting sequence data.

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Research Organization:: Lawrence Berkeley National Laboratory-National Energy Research Scientific Computing Center (NERSC)

Sponsoring Organization:: USDOE Office of Science (SC)

DOE Contract Number:: AC02-05CH11231

OSTI ID:: 1546616

Country of Publication:: United States

Language:: English

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