Methods for detection and quantification of analytes in complex mixtures
View Patent ↗The invention provides a diverse population of uniquely labeled probes, containing target specific nucleic acid probes each attached to a unique label bound to a nucleic acid. Also provided is a method of detecting a nucleic acid analyte. The method consists of (a) contacting a mixture of nucleic acid analytes under conditions sufficient for hybridization with a plurality of target specific nucleic acid probes each having a different specifier; (b) contacting the mixture under conditions sufficient for hybridization with a corresponding plurality of antigenedigits each having a unique label, the plurality of anti-genedigits having a diversity sufficient to uniquely hybridize to genedigits within the specifiers, and (c) uniquely detecting a hybridized complex between one or more analytes in the mixture, a target specific probe, and an anti-genedigit.
1. A probe comprising:
(a) a region that selectively binds to a target nucleic acid analyte,
(b) a region comprising a plurality of label-attachment positions linked together, each label-attachment position being of a predetermined nucleotide sequence, wherein each label-attachment position is attached to a corresponding predetermined label monomer, wherein each label monomer is spatially-separable for visualization and wherein each label monomer comprises a fluorescent moiety; and
(c) a biotin tag;
wherein said plurality of label-attachment positions is at least four label-attachment positions and at least 4 corresponding label monomers.
2. The probe of claim 1 , wherein each label monomer is non-covalently attached to the corresponding label-attachment position.
3. The probe of claim 1 , wherein said plurality of label-attachment positions is at least five label-attachment positions and at least 5 corresponding label monomers.
4. The probe of claim 1 , wherein the plurality of label-attachment positions are in a predetermined order within the probe.
5. The probe of claim 1 , wherein at least two of the label-attachment positions have different nucleotide sequences.
6. The probe of claim 1 , wherein each label-attachment position has a different nucleotide sequence.
7. The probe of claim 6 , wherein nucleotide sequence of each label-attachment position differs by at least two nucleotides.
8. The probe of claim 1 , wherein at least two of the label-attachment positions comprise a repeated core element.
9. The probe of claim 1 , wherein the nucleotide sequence of each label-attachment position comprises at least one isocytosine or isoguanine nucleotide.
10. The probe of claim 1 , wherein the plurality of label-attachment positions are directly linked.
11. The probe of claim 1 , wherein the plurality of label-attachment positions are linked via intervening or adapting sequences.