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Bromodeoxyuridine (BrdU) Labeling and Subsequent Fluorescence Activated Cell Sorting for Culture-independent Identification of Dissolved Organic Carbon-degrading Bacterioplankton
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Biologia
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JoVE Journal Biologia
Bromodeoxyuridine (BrdU) Labeling and Subsequent Fluorescence Activated Cell Sorting for Culture-independent Identification of Dissolved Organic Carbon-degrading Bacterioplankton

Bromodeoxyuridine (BrdU) Labeling and Subsequent Fluorescence Activated Cell Sorting for Culture-independent Identification of Dissolved Organic Carbon-degrading Bacterioplankton

DOI:
10.3791/2855-v

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12:46 min

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September 10, 2011

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, , , ,
1Biological Sciences,Kent State University, 2Marine Sciences,University of Georgia (UGA)

Capitoli

  • 00:05Titolo
  • 02:12Water Sample Processing
  • 03:28Microcosm Establishment and Incubation
  • 05:13In situ Immunodetection for BrdU Incorporation and FACS
  • 08:43Filter PCR Amplification of 16S rRNA Genes
  • 10:42Representative Results
  • 12:05Conclusion

Summary

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Environmental bacterioplankton are incubated with a model dissolved organic carbon (DOC) compound and a DNA labeling reagent, bromodeoxyuridine (BrdU). Afterward, DOC-degrading cells are separated from the bulk community based on their elevated BrdU incorporation using fluorescence activated cell sorting (FACS). These cells are then identified by subsequent molecular analyses.

Tags

BromodeoxyuridineBrdU LabelingFluorescence Activated Cell SortingCulture-independent IdentificationDissolved Organic CarbonBacterioplanktonMicrobial DegradationAquatic Environments16S RRNA Gene-based Molecular AnalysisDOC CompoundsMicrocosmsImmunodetectionHI CellsLI Cells

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