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US 10,526,652 B2Granted
Modified nucleotides methods and kits
AssigneePierce Biotechnology, Inc.
InventorsChristopher L. Etienne, Kay K. Opperman, Barbara J. Kaboord, Scott D. Meier, Jean-Samuel Schultz
Life of this patent
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Abstract
Modified nucleotides, and methods to modify nucleotides with a moiety or label, such as biotin, that permits their detection and results in a modified nucleotide, and methods of use of the modified nucleotide in quantitative and qualitative assays.
Claims16
A method of quantitating a nucleic acid or an oligonucleotide comprising:
- a) adding to a nucleic acid or an oligonucleotide, a detectable modified nucleotide of structure II:
- a salt, a conjugate base, a tautomer, or an ionized form thereof, wherein
- Base* is a purine or pyrimidine base;
- R is H, OH, CH3 or a hydroxyl protecting group;
- Alk is a connecting group having the structure -//—(CH2)m—Y—//- wherein Y is a bond or bond forming group selected from
- and
- m is an integer ranging from 3 to 6 inclusive, and wherein the leftmost bond is to Nus and the rightmost bond is to Lnk;
- Lnk is a linking group having the structure
- wherein
- n is an integer ranging from 2 to 48 inclusive;
- A1 is a bond forming group selected from
- A2 is a bond forming group selected from
- A3, when present, is a bond forming group selected from
- and X is a cleavable group that can undergo silicon-carbon cleavage, nucleophilic cleavage, redox cleavage, photochemical cleavage, enzymatic cleavage, or exchange-based cleavage;
- Z is a branching group that contains a crosslinking agent;
- and the leftmost bond is to Alk and the rightmost bond is to Obs;
- and Obs is an observable label;
- and
- b) quantitating the nucleic add based on quantifying the detectable modified nucleotide.
16 more elements
A method for assaying a ribonucleic acid (RNA) analyte, the method comprising:
- a) labeling an RNA with the detectable modified nucleotide of structure (II)
- a salt, a conjugate base, a tautomer, or an ionized form thereof, wherein
- Base* is a purine or pyrimidine base;
- R is H, OH, CH3, or a hydroxyl protecting group;
- Alk is a connecting group having the structure -//—(CH2)m—Y—//- wherein Y is a bond or bond forming group selected from
- and
- m is an integer ranging from 3 to 6 inclusive, and wherein the leftmost bond is to Nus and the rightmost bond is to Lnk;
- Lnk is a linking group having the structure
- wherein
- n is an integer ranging from 2 to 48 inclusive;
- A1 is a bond forming group selected from
- A2 is a bond forming group selected from
- A3, when present, is a bond forming group selected from
- and X is a cleavable group that can undergo silicon-carbon cleavage, nucleophilic cleavage, redox cleavage, photochemical cleavage, enzymatic cleavage, or exchange-based cleavage;
- Z is a branching group that contains a crosslinking agent;
- and the leftmost bond is to Alk and the rightmost bond is to Obs;
- and Obs is an observable label;
- to form a modified RNA probe,
- b) contacting the modified RNA probe with a sample containing the RNA analyte under conditions to hybridize the modified RNA probe with the RNA analyte, and
- c) detecting the RNA analyte hybridized with the modified RNA probe, wherein hybridization and detection of the modified RNA probe assays the RNA analyte.
17 more elements
Dependencies are inferred from the claim text.