Element-tagged olignucleotide gene expression analysis
OwnerOlga Ornatskyinventor
Life of this patent
Estimating the term…
Abstract
Methods and kits for gene expression analysis are disclosed. The methods utilize element-tagged oligonucleotides as probes which are subsequently analyzed by elemental analysis. Also disclosed are methods and kits for the analysis of biological molecules using element labeled supports such as beads, followed by elemental analysis. The elemental analysis can be done using ICP-MS.
Claims57
A method for cellular analysis, comprising:
- (a) providing a cell or a cellular particle;
- (b) fixing and permeabilizing the cell or the cellular particle;
- (c) incubating the cell or the cellular particle in a hybridization solution with a probe specific for a target nucleic acid, the probe labeled with a unique element tag such that one type of said probe labeled with one type of said tag is distinguishable from any other type of said probe labeled with a different type of said tag by elemental analysis;
- (d) separating unhybridized probe from probe hybridized to the target nucleic acid by stringent washing conditions;
- and
- (e) analyzing the cell or cellular particle by elemental analysis to identify the probe and quantify the probe bound to the target nucleic acid.
The method of example 1 further comprising simultaneous analysis of surface and/or intracellular small molecules, selected from the group consisting of vitamins, hormones, haptens and nucleosides.
A method for homogeneous analysis of biological molecules in solution, comprising:
- (a) incubating biological molecules with affinity reagents labeled with element tags and uniquely tagged supports such that one type of said supports labeled with one type of said tags is distinguishable from any other type of said support labeled with a different type of said tags by elemental analysis, under conditions to enable the affinity reagents to bind with the biological molecules;
- (b) separating the supports with bound biological molecules from unbound supports;
- and
- (c) measuring the bound supports by particle elemental analysis wherein the supports are dispersed in a liquid to measure quantitatively the atomic and isotopic composition of individual supports, thereby detecting the types and the numbers of biological molecules attached to said supports.
A kit for the detection and measurement of an element in a sample, where the measured element is an element tag attached to a specific probe complementary to a nucleic acid of interest, comprising:
- (a) an element tag for directly tagging a complementary probe;
- and
- (b) a complementary probe.
A kit for the detection and measurement of an element in a sample, where the measured element is an element tag attached to a specific probe complementary to a nucleic acid of interest, comprising:
- (a) a complementary probe tagged with an element tag.
A kit for the detection and measurement of an element, where the measured element is an element tag attached to oligo(dT)n and elements of uniquely labeled supports attached to complimentary probe, comprising:
- (a) an element tag for directly tagging oligo(dT)n;
- (b) oligo(dT)n;
- (c) at least one uniquely labeled support;
- and
- (d) a multitude of complementary probes.
A kit for the detection and measurement of an element, where the measured element is an element tag attached to oligo(dT)n and elements of uniquely labeled supports attached to complimentary probes, comprising:
- (a) an element tag for directly tagging oligo(dT)n;
- (b) oligo(dT)n;
- and
- (c) a multitude of complementary probes attached to at least one uniquely labeled supports.
A kit for the detection and measurement of an element, where the measured element is an element tag attached to oligo(dT)n and elements of uniquely labeled supports attached to complimentary probes, comprising:
- (a) an element tag labeled oligo(dT)n;
- and
- (b) a multitude of complementary probes attached to at least one uniquely labeled supports.
Dependencies are inferred from the claim text.