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Targeted genomics of flow cytometrically sorted cultured and uncultured microbial groups
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Mazard, Sophie, Ostrowski, Martin, Holland, Ross, Zubkov, Mikhail V. and Scanlan, David J. (2014) Targeted genomics of flow cytometrically sorted cultured and uncultured microbial groups. In: Paulsen, Ian T. and Holmes, Andrew J., (eds.) Environmental Microbiology. Methods in Molecular Biology, Volume 1096 . Humana Press, pp. 203-212. ISBN 9781627037112
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Official URL: http://dx.doi.org/10.1007/978-1-62703-712-9_16
Abstract
High throughput sequencing of genetic material recovered from environmental samples (i.e., metagenomics) is becoming the method of choice for either medical or environmental genomic studies. However, the large amount of data and complexity of the sequenced “biomes” present challenges for teasing meaningful results out of the mass. Here, we describe a targeted genomic pipeline which uses fluorescence-activated cell sorting (FACS) in combination with multiple displacement amplification (MDA) of nucleic acids that allows to dissect a complex system into its component parts to facilitate high-quality single-cell, or targeted population, genomic reconstructions of microbial communities. This pipeline is presented with methods for collecting, concentrating, and preserving cells from aquatic and marine environments suitable for flow cytometric processing at a later date.
Item Type: | Book Item | ||||
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
Series Name: | Methods in Molecular Biology | ||||
Publisher: | Humana Press | ||||
ISBN: | 9781627037112 | ||||
ISSN: | 1064-3745 | ||||
Book Title: | Environmental Microbiology | ||||
Editor: | Paulsen, Ian T. and Holmes, Andrew J. | ||||
Official Date: | 15 January 2014 | ||||
Dates: |
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Volume: | Volume 1096 | ||||
Page Range: | pp. 203-212 | ||||
DOI: | 10.1007/978-1-62703-712-9_16 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access |
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