High-throughput isotopic analysis of RNA microarrays to quantify microbial resource use.

ABSTRACT: Most microorganisms remain uncultivated, and typically their ecological roles must be inferred from diversity and genomic studies. To directly measure functional roles of uncultivated microbes, we developed Chip-stable isotope probing (SIP), a high-sensitivity, high-throughput SIP method performed on a phylogenetic microarray (chip). This approach consists of microbial community incubations with isotopically labeled substrates, hybridization of the extracted community rRNA to a microarray and measurement of isotope incorporation--and therefore substrate use--by secondary ion mass spectrometer imaging (NanoSIMS). Laboratory experiments demonstrated that Chip-SIP can detect isotopic enrichment of 0.5 atom % (13)C and 0.1 atom % (15)N, thus permitting experiments with short incubation times a

SUBMITTER: Mayali X 

PROVIDER: S-EPMC3358021 | biostudies-literature | 2012 Jun

REPOSITORIES: biostudies-literature