The present disclosure relates in some aspects to proteins and nucleic acids for probing analytes in a biological sample. In some embodiments, disclosed herein is a method in which detection probes (e.g., fluorescently labeled detection probes) may comprise a sequence of nucleobases on a synthetic backbone, the detection probes therefore generating a greater signal than reference detection probes having the same sequence of nucleobases on a sugar-phosphate backbone.

Methods, compositions and kits for capturing, detecting and quantifying mature small RNAs are provided herein. Embodiments of the methods comprise ligating 5′ and 3′ ligation adaptors to the 5′ and 3′ ends of the mature small RNAs, respectively, in the presence of 5′ and 3′ semi-degenerate ligation splints to generate a ligation product. Other embodiments comprise reverse transcribing polyadenylated mature small RNA with a universal reverse transcription primer and ligating an adaptor to the 3′ end of the cDNA in the presence of a semi-degenerate ligation splint to generate a cDNA ligation product.

The invention relates to a method for the detection of natural or modified nucleic acids by their sequence specific hybridization with charge-modified oligonucleotide probes having charge-modifying groups attached to their backbones. The charge-modifying groups partially or fully neutralize the net negative charge of the backbone of the oligonucleotide probes or render them with a net positive charge. The charge-modified oligonucleotide probes may or may not be labeled, for example, with fluorescent, visible or near-infrared dye, with radioactive or stable isotopes, or with high specific affinity binding groups. The charge-modified oligonucleotide probes facilitate the separation of their hybrids with the targeted nucleic acids from the unhybridized probes or from any other components of the analyzed sample. They also allow for the modification and optimization of the properties of the hybrids with the targeted nucleic acids, such as melting temperature, chromatographic properties and off-target specificity.

The invention relates to a method for the detection of natural or modified nucleic acids by their sequence specific hybridization with charge-modified oligonucleotide probes having charge-modifying groups attached to their backbones. The charge-modifying groups partially or fully neutralize the net negative charge of the backbone of the oligonucleotide probes or render them with a net positive charge. The charge-modified oligonucleotide probes may or may not be labeled, for example, with fluorescent, visible or near-infrared dye, with radioactive or stable isotopes, or with high specific affinity binding groups. The charge-modified oligonucleotide probes facilitate the separation of their hybrids with the targeted nucleic acids from the unhybridized probes or from any other components of the analyzed sample. They also allow for the modification and optimization of the properties of the hybrids with the targeted nucleic acids, such as melting temperature, chromatographic properties and off-target specificity.

A non-specific-binding inhibitor enables, in detection of a target nucleic acid by hybridization, effective inhibition of cross-hybridization between the target nucleic acid and complementary strands of sequences similar thereto, which non-specific-binding inhibitor has a stable quality among production lots, and a hybridization method for nucleic acid uses the inhibitor. The non-specific-binding inhibitor for nucleic acid includes a nucleic acid which has a base length of 2 to 11 bases and in which the content of a guanine base(s) and/or methylated guanine base(s) in the entire base sequence is not less than 70%.

A non-specific-binding inhibitor enables, in detection of a target nucleic acid by hybridization, effective inhibition of cross-hybridization between the target nucleic acid and complementary strands of sequences similar thereto, which non-specific-binding inhibitor has a stable quality among production lots, and a hybridization method for nucleic acid uses the inhibitor. The non-specific-binding inhibitor for nucleic acid includes a nucleic acid which has a base length of 2 to 11 bases and in which the content of a guanine base(s) and/or methylated guanine base(s) in the entire base sequence is not less than 70%.

A non-specific-binding inhibitor enables, in detection of a target nucleic acid by hybridization, effective inhibition of cross-hybridization between the target nucleic acid and complementary strands of sequences similar thereto, which non-specific-binding inhibitor has a stable quality among production lots, and a hybridization method for nucleic acid uses the inhibitor. The non-specific-binding inhibitor for nucleic acid includes a nucleic acid which has a base length of 2 to 11 bases and in which the content of a guanine base(s) and/or methylated guanine base(s) in the entire base sequence is not less than 70%.

Provided herein are primers and primer systems having improved specificity and kinetics over existing primers, and methods of use thereof.

Provided herein are primers and primer systems having improved specificity and kinetics over existing primers, and methods of use thereof.

Provided herein are primers and primer systems having improved specificity and kinetics over existing primers, and methods of use thereof.