DNA stable-isotope probing
作者： Josh D Neufeld , Jyotsna Vohra , Marc G Dumont , Tillmann Lueders , Mike Manefield
关键词: Nucleic acid 、 Stable-isotope probing 、 DNA sequencing 、 Biochemistry 、 DNA microarray 、 DNA 、 Biology 、 Transduction (genetics) 、 Recombinant DNA 、 Genomics
摘要: Stable-isotope probing is a method used in microbial ecology that provides means by which specific functional groups of organisms incorporate particular substrates are identified without the prerequisite cultivation. Stable-isotope-labeled carbon (13C) or nitrogen (15N) sources assimilated into biomass environmental samples. Separation and molecular analysis labeled nucleic acids (DNA RNA) reveals phylogenetic information about microorganisms responsible for metabolism substrate. Here, we highlight general guidelines incubating samples with substrate provide detailed protocol separating DNA from unlabeled community DNA. The includes modification existing published methods, maximizes recovery CsCl gradients. separation retrieval fractions can be performed 4-5 days, much time being committed to ultracentrifugation step.
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