US4626501A
Expired / Lapsed
US
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Labeled DNA
Assignee: Genzyme Corp
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Filing: 1983-09-02
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Granted: 1986-12-02
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Expires: 2003-09-02
Abstract
Abstract Polymeric, single-stranded DNA molecule which is labeled with a plurality of labels, the labels being bonded, via linking groups, to adenine and cytosine bases of the DNA molecule in substantially equal numbers, the labels being bonded to essentially none of the guanine or thymine bases of the DNA molecule, the DNA molecule being adapted for use as a probe in a nucleic acid hybridization assay.
Inventors
- 👤 Greg M. Landes
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 11:39
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C07; C07H; C07H21/00; C12Q1/6825; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/535
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
111 words
A polymeric DNA molecule wherein at least some of the adenine and cytosine bases of said DNA molecule are derivatized to their respective etheno analogs, said analogs having the formula ##STR11## wherein n is between 0 and 10, inclusive; each p is between 1 and 10, inclusive; n a is N-1 of adenine or N-3 of cytosine; N b is N-6 of adenine or N-4 of cytosine; B is the remainder of the adenine or cytosine base; W is SO 2 --(CH 2 ) q , q being between 1 and 10, inclusive; and G is X, X being NH--(CH 2 ) r --Z wherein 1≦r≦10, Z being a detectable entity.
Claim 2
Dependent Claim
111 words
A polymeric DNA molecule wherein at least some of the adenine and cytosine bases of said DNA molecule are derivatized to their respective etheno analogs, said analogs having the formula ##STR11## wherein n is between 0 and 10, inclusive; each p is between 1 and 10, inclusive; n a is N-1 of adenine or N-3 of cytosine; N b is N-6 of adenine or N-4 of cytosine; B is the remainder of the adenine or cytosine base; W is SO 2 --(CH 2 ) q , q being between 1 and 10, inclusive; and G is X, X being NH--(CH 2 ) r --Z wherein 1≦r≦10, Z being a detectable entity.
Claim 3
Dependent Claim
27 words
The DNA molecule of claim 1 wherein n is 0, p is 2, and W is SO 2 --(CH 2 ) q , wherein q is 2.
Claim 4
Dependent Claim
28 words
The DNA molecule of claim 1 wherein at least half of the adenine bases and at least half of the cytosine bases of said DNA molecule are derivatized.
Claim 5
Dependent Claim
18 words
The DNA molecule of claim 1 wherein G is X, X comprising one of a specific binding pair.
Claim 6
Dependent Claim
15 words
The DNA molecule of claim 1 wherein G is X, X comprising a detectable enzyme.
Claim 7
Dependent Claim
15 words
The DNA molecule of claim 1 wherein G is X, X comprising a detectable hapten.
Claim 8
Dependent Claim
16 words
The DNA molecule of claim 1 wherein G is X, X comprising a radioactively detectable atom.
Claim 9
Dependent Claim
17 words
The DNA molecule of claim 1 wherein G is X, X comprising a spectro photometrically detectable molecule.
Claim 10
Dependent Claim
15 words
The DNA molecule of claim 1 wherein G is X, X comprising a fluorescent molecule.
Claim 11
Dependent Claim
16 words
The DNA molecule of claim 1 wherein Z is biotin or an avidin-specific binding derivative thereof.
Claim 12
Dependent Claim
15 words
The DNA molecule of claim 10 wherein said specific binding derivative of biotin is ##STR12##
Claim 13
Dependent Claim
22 words
The DNA molecule of claim 4 wherein said one of a specific binding pair is one of a specific binding immunological pair.
Claim 14
Dependent Claim
21 words
The DNA molecule of claim 12 wherein said one of a specific binding immunological pair is one of an antibody/antigen pair.
Claim 15
Dependent Claim
12 words
The DNA molecule of claim 1 wherein Z is a fluorescent molecule.
Claim 16
Dependent Claim
14 words
The DNA molecule of claim 1 wherein Z is a spectro photometrically detectable molecule.
Claim 17
Dependent Claim
13 words
The DNA molecule of claim 1 wherein Z is a radioactively detectable atom.
Claim 18
Dependent Claim
18 words
The DNA molecule of claim 1 wherein said etheno analogs of said DNA molecule have the formula: ##STR13##
Claim 19
Dependent Claim
18 words
The DNA molecule of claim 1 wherein said etheno analogs of said DNA molecule have the formula: ##STR14##
Claim 20
Dependent Claim
26 words
The DNA molecule of claim 1 wherein said etheno analogs of said DNA molecule have the formula: ##STR15## wherein r is between 1 and 10, inclusive.
Claim 21
Dependent Claim
26 words
The DNA molecule of claim 1 wherein said etheno analogs of said DNA molecule have the formula: ##STR16## wherein r is between 1 and 10, inclusive.
Claim 22
Dependent Claim
61 words
A polymeric DNA molecule wherein at least some of the adenine and cytosine basis of said DNA molecule are derivatized to their respective etheno analogs having the formula: ##STR17## wherein N a is N-1 of adenine or N-3 of cytosine; N b is N-6 of adenine or N-4 of cytosine; and B is the remainder of said adenine or cytosine base.
Claim 23
Dependent Claim
61 words
A polymeric DNA molecule wherein at least some of the adenine and cytosine basis of said DNA molecule are derivatized to their respective etheno analogs having the formula: ##STR17## wherein N a is N-1 of adenine or N-3 of cytosine; N b is N-6 of adenine or N-4 of cytosine; and B is the remainder of said adenine or cytosine base.
Claim 24
Independent Claim
188 words
A method of making a probe for use in a nucleic acid hybridization assay for a specific nucleic acid sequence, said method comprising providing a single-stranded polymeric DNA molecule capable of specifically hybridizing with said nucleic acid sequence, reacting said DNA molecule with a compound of the formula ##STR18## wherein V is a halogen; n is between 0 and 10, inclusive; and each p and q, independently, is between 1 and 10, inclusive, whereby at least some of the adenine and cytosine bases of said DNA molecule are derivatized to their respective etheno analogs, said analogs having the formula ##STR19## wherein n is between 0 and 10, inclusive; each p and q, independently, is between 1 and 10, inclusive; N a is N-1 of adenine or N-3 of cytosine; N b is N-6 of adenine or N-4 of cytosine; and B is the remainder of the adenine or cytosine base, performing one or more steps to substitute, for ##STR20## on said analogs, a substituent comprising a label which either is capable of being detected or is capable of selectively bonding to an indicator to form a detectable complex.
Claim 25
Independent Claim
188 words
A method of making a probe for use in a nucleic acid hybridization assay for a specific nucleic acid sequence, said method comprising providing a single-stranded polymeric DNA molecule capable of specifically hybridizing with said nucleic acid sequence, reacting said DNA molecule with a compound of the formula ##STR18## wherein V is a halogen; n is between 0 and 10, inclusive; and each p and q, independently, is between 1 and 10, inclusive, whereby at least some of the adenine and cytosine bases of said DNA molecule are derivatized to their respective etheno analogs, said analogs having the formula ##STR19## wherein n is between 0 and 10, inclusive; each p and q, independently, is between 1 and 10, inclusive; N a is N-1 of adenine or N-3 of cytosine; N b is N-6 of adenine or N-4 of cytosine; and B is the remainder of the adenine or cytosine base, performing one or more steps to substitute, for ##STR20## on said analogs, a substituent comprising a label which either is capable of being detected or is capable of selectively bonding to an indicator to form a detectable complex.
Claim 26
Dependent Claim
170 words
A polymeric, single-stranded DNA molecule adapted for use as a probe in a nucleic acid hybridization assay for a specific nucleic acid sequence, said molecule comprising a first portion capable of specifically hybridizing with said specific nucleic acid sequence, and a second tail portion not capable of so hybridizing, said tail portion being labeled with a plurality of labels and said first portion being substantially unlabeled wherein said tail contains adenine or cytosine bases, and is substantially free of guanine and thymine bases, and wherein at least some of said adenine or cytosine bases have bonded to them a substituent of the formula ##STR21## wherein n is between 0 and 10, inclusive; each p and q, independently, is between 1 and 10, inclusive; N a is N-1 of adenine or N-3 of cytosine; N b is N-6 of adenine or N-4 of cytosine; B is the remainder of the adenine or cytosine base; and X is NH--(CH 2 ) r --Z, wherein 1≦r≦10 and Z is a detectable chemical entity.
Claim 27
Dependent Claim
170 words
A polymeric, single-stranded DNA molecule adapted for use as a probe in a nucleic acid hybridization assay for a specific nucleic acid sequence, said molecule comprising a first portion capable of specifically hybridizing with said specific nucleic acid sequence, and a second tail portion not capable of so hybridizing, said tail portion being labeled with a plurality of labels and said first portion being substantially unlabeled wherein said tail contains adenine or cytosine bases, and is substantially free of guanine and thymine bases, and wherein at least some of said adenine or cytosine bases have bonded to them a substituent of the formula ##STR21## wherein n is between 0 and 10, inclusive; each p and q, independently, is between 1 and 10, inclusive; N a is N-1 of adenine or N-3 of cytosine; N b is N-6 of adenine or N-4 of cytosine; B is the remainder of the adenine or cytosine base; and X is NH--(CH 2 ) r --Z, wherein 1≦r≦10 and Z is a detectable chemical entity.
Claim 28
Independent Claim
130 words
A method of making a probe for use in a nucleic acid hybridization assay for a specific nucleic acid sequence, said method comprising providing a single-stranded polymeric DNA molecule capable of specifically hybridizing with said nucleic acid sequence, reacting said DNA molecule with a compound of the formula ##STR22## wherein V is a halogen; n is between 0 and 10, inclusive; and each p and q, independently, is between 1 and 10, inclusive, whereby at least some of the adenine and cytosine bases of said DNA molecule are derivatized to etheno analogs, performing one or more steps to substitute, for ##STR23## on said analogs, a substituent comprising a label which either is capable of being detected or is capable of selectively bonding to an indicator to form a detectable complex.
Claim 29
Independent Claim
130 words
A method of making a probe for use in a nucleic acid hybridization assay for a specific nucleic acid sequence, said method comprising providing a single-stranded polymeric DNA molecule capable of specifically hybridizing with said nucleic acid sequence, reacting said DNA molecule with a compound of the formula ##STR22## wherein V is a halogen; n is between 0 and 10, inclusive; and each p and q, independently, is between 1 and 10, inclusive, whereby at least some of the adenine and cytosine bases of said DNA molecule are derivatized to etheno analogs, performing one or more steps to substitute, for ##STR23## on said analogs, a substituent comprising a label which either is capable of being detected or is capable of selectively bonding to an indicator to form a detectable complex.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-12-01 | 1990-06-02 | 1990-12-02 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-12-01 | 1994-06-02 | 1994-12-02 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-12-01 | 1998-06-02 | 1998-12-02 | $7700.00 | Window Closed / Lapsed |