Reagents for the preparation of 5'-tagged oligonucleotides
First Claim
1. A nonnucleoside-containing chemical reagent having the formula
wherein Q is a phosphorus group suitable for attachment to the 5'"'"'-hydroxyl group of an oligonucleotide selected from the group of phosphorus groups consisting of ##STR13## and salts thereof;
wherein R2 and R3 are independently selected from the group consisting of C3 -C10 branched alkyl and C1 -C10 unbranched alkyl;
wherein R4 is selected from the group consisting of --(CRR1)m (A)n (CRR1)m --, ##STR14## wherein A is selected from the group consisting of ##STR15## wherein R and R1 are independently selected from the group consisting of H, C3 -C10 branched alkyl, C1 -C10 unbranched alkyl, C6 -C10 aryl, C7 -C12 alkaryl, and C7 -C12 aralkyl;
wherein m=1-6, n=0-1, p=1-10, and q=0-10, provided that 2m+n≦
12 and further provided that 2≦
n+p+q ≦
13;
wherein X is selected from the group consisting of --F, --Cl, --Br, --I, imidazol-1-yl, 1,2,4-triazol-1yl, and 1-hydroxybenzotriazol-O-yl;
wherein Y is selected from the group consisting of 4-Cl-C6 H4 -O-, 2--Cl--C6 H4 --O--, 4--NO2 --C6 H4 --O--, 4NO2 --C6l H4 CH2 CH2 --O--, 2,4--NO2 --C6 H3 CH2 CH2 --O--, 2,4-Cl2 --C6 H3 --O--, 2,3--Cl2 --C6 H3 --O--, NCCH2 CH2 O--, NCCH2 C(CH3)2 --O--, CH3 O--, (Z)3 CCH2 --O--, (Z)3 CC(CH3)2 --O--, R'"'"'S--, R'"'"'SCH2 CH2 --O--, R'"'"'SO2 CH2 CH2 --O--, and R'"'"'NH--;
wherein Z is selected from the group consisting of Cl, Br, and I;
wherein R'"'"' is selected from the group consisting of H, C3 -C10 branched alkyl, C1 -C10 unbranched alkyl, C6 -C10 aryl, C7 -C12 alkaryl, and C7 -C12 aralkyl;
wherein the --O-- in --O--reporter was formerly part of a hydroxyl group and is covalently linked to said Q; and
wherein the --O--reporter is either (i) a fluorescent reporter selected from the group of fluorescent dyes consisting of fluoresceins, benzamidizoles, ethidium,s propidiums, anthracyclines, mithramycins, actinomycins, merocyanines, coumarins, pyrenes, chrysenes, stilbenes, anthracenes, naphthalenes, salicylic acids, benz-2-oxadiazoles, and fluorescamine, or (ii) a protected from of one of said fluorescent reporters.
2 Assignments
0 Petitions
Accused Products
Abstract
Fluorescent reagents possessing an activated phosphate for the convenient covalent coupling to the 5'"'"'-hydroxyl of oligonucleotides. A class of 5'"'"'-fluorescence-tagged oligonucleptides is also disclosed.
-
Citations
6 Claims
- 1. A nonnucleoside-containing chemical reagent having the formula
- space="preserve" listing-type="equation">Q--O--reporter
wherein Q is a phosphorus group suitable for attachment to the 5'"'"'-hydroxyl group of an oligonucleotide selected from the group of phosphorus groups consisting of ##STR13## and salts thereof;
wherein R2 and R3 are independently selected from the group consisting of C3 -C10 branched alkyl and C1 -C10 unbranched alkyl;wherein R4 is selected from the group consisting of --(CRR1)m (A)n (CRR1)m --, ##STR14## wherein A is selected from the group consisting of ##STR15## wherein R and R1 are independently selected from the group consisting of H, C3 -C10 branched alkyl, C1 -C10 unbranched alkyl, C6 -C10 aryl, C7 -C12 alkaryl, and C7 -C12 aralkyl; wherein m=1-6, n=0-1, p=1-10, and q=0-10, provided that 2m+n≦
12 and further provided that 2≦
n+p+q ≦
13;wherein X is selected from the group consisting of --F, --Cl, --Br, --I, imidazol-1-yl, 1,2,4-triazol-1yl, and 1-hydroxybenzotriazol-O-yl; wherein Y is selected from the group consisting of 4-Cl-C6 H4 -O-, 2--Cl--C6 H4 --O--, 4--NO2 --C6 H4 --O--, 4NO2 --C6l H4 CH2 CH2 --O--, 2,4--NO2 --C6 H3 CH2 CH2 --O--, 2,4-Cl2 --C6 H3 --O--, 2,3--Cl2 --C6 H3 --O--, NCCH2 CH2 O--, NCCH2 C(CH3)2 --O--, CH3 O--, (Z)3 CCH2 --O--, (Z)3 CC(CH3)2 --O--, R'"'"'S--, R'"'"'SCH2 CH2 --O--, R'"'"'SO2 CH2 CH2 --O--, and R'"'"'NH--; wherein Z is selected from the group consisting of Cl, Br, and I; wherein R'"'"' is selected from the group consisting of H, C3 -C10 branched alkyl, C1 -C10 unbranched alkyl, C6 -C10 aryl, C7 -C12 alkaryl, and C7 -C12 aralkyl; wherein the --O-- in --O--reporter was formerly part of a hydroxyl group and is covalently linked to said Q; and wherein the --O--reporter is either (i) a fluorescent reporter selected from the group of fluorescent dyes consisting of fluoresceins, benzamidizoles, ethidium,s propidiums, anthracyclines, mithramycins, actinomycins, merocyanines, coumarins, pyrenes, chrysenes, stilbenes, anthracenes, naphthalenes, salicylic acids, benz-2-oxadiazoles, and fluorescamine, or (ii) a protected from of one of said fluorescent reporters. - View Dependent Claims (2, 3, 4, 5, 6)
Specification
-
- Resources
-
Current AssigneeNEN Life Science Products Inc (Revvity, Inc.)
-
Original AssigneeE. I. du Pont de Nemours and Company (Corteva, Inc.)
-
InventorsHobbs, Frank W. Jr.
-
Primary Examiner(s)Brown, Johnnie R.
-
Assistant Examiner(s)Kunz, Gary L.
-
Application NumberUS07/622,265Time in Patent Office1,077 DaysField of Search435/6, 536/27, 536/28, 536/29, 536/25.32, 558/93, 558/95, 558/168, 558/191, 558/192, 558/218, 549/385, 549/223, 549/283, 546/242, 546/25US Class Current546/25CPC Class CodesC07F 9/65586 at least one of the hetero ...C07H 21/00 Compounds containing two or...C12Q 1/686 Polymerase chain reaction [...C12Q 1/6876 Nucleic acid products used ...G01N 33/582 with fluorescent label
