A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:04Title
- 01:07Introduction
- 02:11Sample Incubation and DNA Extraction
- 03:47Gradient Solns for Ultracentrifugation
- 05:26Ultracentrifugation
- 07:49Gradient Fractionation
- 09:46DNA Precipitation
- 11:22Representative Results
- 14:04Conclusion
DNA stable-isotope probing is a cultivation-independent method to identify and characterize active communities of microorganisms that are capable of utilizing specific substrates. Assimilation of substrate enriched in heavy isotope leads to incorporation of labelled atoms into microbial biomass. Density gradient ultracentrifugation retrieves labelled DNA for downstream molecular analyses.
Tags
DNA-SIPStable-isotope ProbingMicroorganismsCarbon SubstratesNutrientsCellular BiomassCultivation-independent TechniqueMetabolic FunctionTerrestrial EnvironmentsAquatic EnvironmentsStable-isotope Labelled CompoundsNucleic AcidDensity Gradient UltracentrifugationGradient FractionationDNA PurificationCesium ChlorideMolecular CharacterizationFingerprintingMicroarraysClone LibrariesMetagenomics
