Quantitative substrate utilization in microbial ecology using stable isotope probing
AssigneeArizona Board of Regents On Behalf of Northern Arizona University
InventorsBruce Hungate, Egbert Schwartz, Rebecca Mau, Jane Marks
Life of this patent
Estimating the term…
Abstract
Methods modifying conventional SIP so that isotopic incorporation into the genomes of individual microbial taxa can be quantified are described. Further, methods to quantify the baseline densities of the DNA of individual microbial taxa without exposure to isotope tracers and then to quantify the change in DNA density of each taxon caused by isotope incorporation are described. The distribution of DNA of each taxon along a density gradient reflects the influence of isotope incorporation only, without reflecting the guanine-plus-cytosine content.
Claims17
A method to quantify an uptake of a substrate by a microorganism sample, comprising:
- extracting nucleic acids from said microorganism sample both with and without incubation with at least one isotopically labeled substrate, thereby resulting in a labeled sample and an unlabeled sample;
- separating said nucleic acids by density into a plurality of labeled or unlabeled fractions;
- measuring a density of said nucleic acids from each of said labeled or unlabeled fractions;
- measuring a relative abundance of a taxon of a microorganism in each of said labeled or unlabeled fractions;
- quantifying a combined density of said taxon of said microorganism in said plurality of labeled or unlabeled fractions;
- and
- measuring a difference between the density of said taxon of said microorganism in the plurality of labeled fractions and the density of said taxon of said microorganism in the corresponding plurality of unlabeled fractions;
- wherein said difference in density is indicative of said uptake of the substrate by said microorganism.
Dependencies are inferred from the claim text.