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US 2014/0272934 A1Published application
Dendrimeric dye-containing oligonucleotide probes and methods of preparation and uses thereof
AssigneeAgilent Technologies, Inc.
InventorsJeremy Lackey, Siyuan Chen, Maithreyan Srinivasan
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Abstract
The invention provides novel oligonucleotide probes that have dendrimeric dyes useful for detecting and analyzing biological samples, and compositions and methods thereof. The dendrimeric dye-containing oligonucleotide probes are useful for high sensitivity fluorescence in situ hybridization (FISH) of nucleic acids such as DNA and RNA.
Claims20
An oligonucleotide probe, comprising:
- a nucleic acid sequence capable of hybridization with a target nucleic acid of interest;
- and
- a primer covalently attached to the oligonucleotide, wherein the primer is covalently attached to a dendrimeric group comprising two or more fluorescent moieties.
A method for generating oligonucleotide probes, comprising:
- generating a library of oligonucleotides;
- providing a plurality of primers each comprising an alkyne moiety;
- forming a library of amplicons wherein each amplicon comprises a member of the library of oligonucleotides and a primer from the plurality of primers;
- providing a dendrimeric group comprising two or more fluorescent moieties and comprises an azide moiety;
- and
- reacting the amplicons with the dendrimeric group by reacting the alkyne moiety with the azide moiety thereby generating a library of oligonucleotide probes.
3 more elements
A method for detecting or analyzing a target nucleic acid of interest, comprising:
- providing an oligonucleotide probe comprising a nucleic acid sequence capable of hybridization with a target nucleic acid of interest and a primer covalently attached a first reactive moiety;
- contacting the oligonucleotide probe with a sample to be analyzed for the presence of the target nucleic acid of interest under a condition such that, if the target nucleic acid of interest is present, a hybridization complex is formed between the oligonucleotide probe and the target nucleic acid of interest;
- and
- reacting the contacted sample with a dedrimeric imaging agent comprising two or more fluorescent moieties and a second reactive moiety, whereby the second reactive moiety reacts with the first reactive moiety to form a covalent bond;
- and
- imaging the two or more fluorescent moieties to detect or analyze the presence of the target nucleic acid of interest.
3 more elements
Dependencies are inferred from the claim text.