US 6,235,483 B1 · Methods and kits for indirect labeling of nucleic acids

Abstract

Methods and kits for labeling nucleic acids are provided. In the subject methods, an oligonucleotide tagged nucleic acid comprising an oligonucleotide tag is first generated. The oligonucleotide tagged nucleic acid is then contacted under hybridization conditions with a labeled oligonucleotide complementary to the oligonucleotide tag, yielding a labeled nucleic acid. The kits of the subject invention at least include a primer for use in enzymatically generating an oligonucleotide tagged target nucleic acid, where the primer generally at least includes an oligo dT region and the oligonucleotide tag, and a labeled oligonucleotide complementary to the oligonucleotide tag. The subject methods and kits find use in a variety of applications, and are particularly suited for use in gene expression analysis applications.

Claims12

Dependent claims

A method of detecting the presence of a nucleic acid analyte in a target sample, said method comprising:

(a) enzymatically generating an oligonucleotide tagged target nucleic acid from said nucleic acid analyte, wherein said oligonucleotide tagged target nucleic acid comprises an oligonucleotide tag;
and
(b) producing a hybridized complex that comprises:
(i) said oligonucleotide tagged target nucleic acid;
(ii) a labeled oligonucleotide complementary to said oligonucleotide tag;
and (iii) a probe nucleic acid, wherein said labeled oligonucleotide is hybridized to said oligonucleotide tag and said probe nucleic acid is hybridized to a non-tag region of said oligonucleotide tagged target nucleic acid;
(c) detecting the presence of said hybridized complex by detecting the presence of said labeled oligonucleotide;
and
(d) relating the presence of said hybridized complex to the presence of said nucleic acid analyte in said sample;
whereby the presence of said nucleic acid analyte in said sample is detected.

7 more elements

Dependencies are inferred from the claim text.

Methods and kits for indirect labeling of nucleic acids

Life of this patent

Methods and kits for indirect labeling of nucleic acids

Life of this patent

Abstract

Claims12