Suppression of non-specific signals by exonucleases in fish experiment
AssigneeCalifornia Institute of Technology
InventorsLong CAI, Chee Huat (Linus) ENG
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Abstract
Disclosed herein is a composition for nuclease treatment of hybridized probes in experiments to profile and analyze biological samples. Also disclosed herein, is a kit for nuclease treatment of hybridized probes in experiments to profile and analyze biological samples. Also disclosed herein, is a method for nuclease treatment of hybridized probes in experiments to profile and analyze biological samples.
Claims33
A composition to reduce the background signal of probes hybridized to targets, wherein the composition comprises:
- (i) one or more primary probes capable of binding one or more targets, wherein each primary probe comprises one or more secondary probe binding sites and/or one or more readout probe binding sites;
- (ii) optionally, one or more secondary probes, each capable of binding the primary probe or one or more targets, wherein each secondary probe comprises one or more tertiary probe binding sites or one or more readout probe binding sites;
- (iii) optionally, one or more tertiary probes, each capable of binding to the secondary probe or one or more targets, wherein each tertiary probe comprises one or more quaternary probe binding sites or one or more readout probe binding sites;
- (iv) optionally, one or more quaternary probes, each capable of binding to the tertiary probe or one or more targets, wherein each quaternary probe comprises one or more readout probe binding sites;
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- (v) one or more readout probes capable of binding to a readout probe binding site on the one or more primary, secondary, tertiary, or quaternary probes and capable of being detected.
- (vi) one or more exonucleases each capable of removing one or more probes that bind to targets, primary, secondary, tertiary, or quaternary probes.
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A method to reduce the background signal of probes hybridized to targets, comprising steps of:
- (i) contacting a sample with one or more primary probes that bind one or more targets, wherein each primary probe hybridizes to a target, wherein each primary probe comprises one or more secondary probe binding sites and/or one or more readout probe binding sites;
- (ii) optionally, hybridizing one or more secondary probes to the primary probes or to one or more targets;
- wherein each secondary probe comprises one or more tertiary probe binding sites or one or more readout probe binding sites;
- (iii) optionally, hybridizing one or more tertiary probes to at least one secondary probe or one or more targets, wherein each tertiary probe comprises one or more quaternary probe binding sites or one or more readout probe binding sites;
- (iv) optionally, hybridizing one or more quaternary probes to at least one tertiary probe or one or more targets, wherein each quaternary probe comprises one or more readout probe binding sites;
- (v) hybridizing readout probes capable of detection to the one or more readout probe binding sites;
- (vi) contacting the sample one or more exonucleases each capable of removing probes that bind to targets, primary, secondary, tertiary, or quaternary probes after any steps (i)-(v) to reduce the background signal of probes bound to targets.
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Dependencies are inferred from the claim text.