High-sensitivity stable-isotope probing by a quantitative terminal restriction fragment length polymorphism protocol.

ABSTRACT: Stable-isotope probing (SIP) has proved a valuable cultivation-independent tool for linking specific microbial populations to selected functions in various natural and engineered systems. However, application of SIP to microbial populations with relatively minor buoyant density increases, such as populations that utilize compounds as a nitrogen source, results in reduced resolution of labeled populations. We therefore developed a tandem quantitative PCR (qPCR)-TRFLP (terminal restriction fragment length polymorphism) protocol that improves resolution of detection by quantifying specific taxonomic groups in gradient fractions. This method combines well-controlled amplification with TRFLP analysis to quantify relative taxon abundance in amplicon pools of FAM-labeled PCR products, using the i

SUBMITTER: Andeer P 

PROVIDER: S-EPMC3255627 | biostudies-literature | 2012 Jan

REPOSITORIES: biostudies-literature