Detection of Nucleic Acids and Proteins
First Claim
1. A method of detecting a nucleic acid and protein, which comprises:
- providing a sample comprising or suspected of comprising a target nucleic acid and a target protein;
incubating at least two label extender probes each comprising a different L-1 sequence, an antibody specific for the target protein, and at least two label probe systems with the sample comprising or suspected of comprising the target nucleic acid and the target protein, wherein the antibody comprises a pre-amplifier probe, and wherein the at least two label probe systems each comprise a detectably different label; and
detecting the detectably different labels in the sample.
3 Assignments
0 Petitions
Accused Products
Abstract
Methods of detecting various types of nucleic acids, including methods of detecting two or more nucleic acids in multiplex branched-chain DNA assays, are provided. Detection assays may be conducted at least in vitro, in cellulo, and in situ. Nucleic acids which are optionally captured on a solid support are detected, for example, through cooperative hybridization events that result in specific association of a label probe system with the nucleic acids. Various label probe system embodiments are provided. Embodiments are directed to concurrent detection of one or more nucleic acids and one or more proteins. Embodiments also are directed to determining the methylation state of a target sequence. Other embodiments are directed to detection of one or more proteins using DNA barcodes. Compositions, kits, and systems related to the methods are also described.
75 Citations
24 Claims
-
1. A method of detecting a nucleic acid and protein, which comprises:
-
providing a sample comprising or suspected of comprising a target nucleic acid and a target protein; incubating at least two label extender probes each comprising a different L-1 sequence, an antibody specific for the target protein, and at least two label probe systems with the sample comprising or suspected of comprising the target nucleic acid and the target protein, wherein the antibody comprises a pre-amplifier probe, and wherein the at least two label probe systems each comprise a detectably different label; and detecting the detectably different labels in the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
-
-
11. A method of detecting a protein, which comprises:
-
providing a sample comprising or suspected of comprising a target protein; incubating an antibody with the sample, wherein the antibody comprises at least one pre-amplifier probe sequence; incubating at least one label probe system with the sample; and detecting whether the label probe system is associated with the sample. - View Dependent Claims (12, 13, 14)
-
-
15. A method of detecting a protein, which comprises:
-
providing a sample comprising or suspected of comprising a target protein; incubating an antibody with the sample, wherein the antibody comprises at least one barcode probe sequence; isolating the antibodies which bind to the sample; and identifying the at least one barcode probe sequence which specifically bound to the sample, thereby detecting the protein in the sample. - View Dependent Claims (16, 17)
-
-
18. A method of determining the methylation state of a nucleic acid sequence, which comprises:
-
providing a sample comprising or suspected of comprising a target nucleic acid sequence; incubating at least two pairs of label extender probes each comprising a different L-1 sequence, at least one pre-amplifier comprising a sequence which is complementary to the target sequence in a region where the methylation status is unknown, and at least three label probe systems with the sample, wherein the at least three label probe systems each comprise a detectably different label; and detecting the detectably different labels in the sample. - View Dependent Claims (19, 20, 21, 22, 23, 24)
-
Specification