Back to results
US 2011/0212850 A1Published application
Using phylogenetic probes for quantification of stable isotope labeling and microbial community analysis
AssigneeThe Regents of the University of California
InventorsEoin L. Brodie, Todd Z. DeSantis, Ulas Karaoz, Gary L. Andersen
Life of this patent
Estimating the term…
Abstract
Herein is described methods for a high-sensitivity means to measure the incorporation of stable isotope labeled substrates into RNA following stable isotope probing experiments (SIP). RNA is hybridized to a set of probes such as phylogenetic microarrays and isotope incorporation is quantified such as by secondary ion mass spectrometer imaging (NanoSIMS).
Claims20
A method for determination of stable isotope incorporation in a organism or a community of organisms comprising the steps of:
- a) supplying an organism or said community of organisms with a stable isotope labeled substrate for a defined period of time;
- b) extracting RNA from the organisms;
- c) fragmenting said RNA;
- d) labeling a fraction the fragmented RNA with a detectable label;
- e) hybridizing the labeled RNA to a set of oligonucleotide probes;
- f) detecting hybridization signal strength of labeled RNA hybridized to any of the oligonucleotide probes and identifying and selecting the hybridized oligonucleotide probes as a responsive set of probes;
- g) hybridizing a fraction of unlabeled RNA to a second set of oligonucleotide probes comprising the responsive set of probes;
- h) detecting the unlabeled RNA hybridized to the responsive set of probes to determine the stable isotope incorporation into the organism using spectrometry or spectroscopy.
5 more elements
Dependencies are inferred from the claim text.