Patent EP0227976A2 successfully refreshed from Google Patents.
EP0227976A2
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Molecules useful for detecting nucleic acid sequences
Assignee: Meiogenics Inc
•
Filing: 1986-12-04
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Granted: —
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Expires: 2006-12-04
Abstract
Abstract A synthetic, non-naturally occurring molecule having the structure: wherein NA 1 and NA 2 are noncomplementary nucleic acid sequences; wherein ---S--- is a scissile linkage; and wherein n is an integer from 1 to 1000. Variations of this molecule and methods for using the molecules for detecting nucleic acid sequences are provided.
Inventors
- 👤 Peter Duck
- 👤 Robert Bender
- 👤 William Crosby
- 👤 John G. Robertson
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-27 22:30
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/70; C12Q1/701; C07; C07H; C07H21/00; C12Q1/68; C12Q1/6813; C12Q1/6816; C12Q1/6823; C12Q1/6834; C12Q1/6876
Claims Analysis
Total Claims: 66
Claim 1
Independent Claim
35 words
A synthetic, non-naturally occurring molecule having the structure: wherein NA 1 and NA 2 are noncomplementary nucleic acid sequences; wherein ---S--- is a scissile linkage; and wherein n is an integer from 1 to 1,000.
Claim 2
Dependent Claim
35 words
A synthetic, non-naturally occurring molecule having the structure: wherein NA 1 and NA 2 are noncomplementary nucleic acid sequences; wherein ---S--- is a scissile linkage; and wherein n is an integer from 1 to 1,000.
Claim 3
Dependent Claim
12 words
The molecule of claim 1, wherein the dashed lines represent chemical bonds.
Claim 4
Dependent Claim
12 words
The molecule of claim 2, wherein the chemical bonds are covalent bonds.
Claim 5
Dependent Claim
12 words
The molecule of claim 2, wherein the chemical bonds are hydrogen bonds.
Claim 6
Dependent Claim
30 words
The molecule of claim 1, wherein the scissile linkage is selected from the group consisting of RNA sequences, DNA sequences, amino acid sequences, abasic nucleic acid sequences or carbohydrate polymers.
Claim 7
Dependent Claim
14 words
The molecule of claim 1, wherein the scissile linkage is a nucleic acid sequence.
Claim 8
Dependent Claim
20 words
The molecule of claim 6, wherein the nucleic acid sequence differs from the sequences of NA1 and NA 2 .
Claim 9
Dependent Claim
10 words
The molecule of claim 1 immobilized on a solid support.
Claim 10
Dependent Claim
14 words
The molecule of claim 1, wherein NA 1 and NA 2 are DNA sequences.
Claim 11
Dependent Claim
14 words
The molecule of claim 1, wherein NA 1 and NA 2 are RNA sequences.
Claim 12
Dependent Claim
25 words
The molecule of claim 1, wherein NA 1 is either an RNA or DNA sequence and NA 2 is either an RNA or DNA sequence.
Claim 13
Dependent Claim
19 words
The molecule of claim 1, wherein NA 1 or NA 2 is not a naturally occurring nucleic acid sequence.
Claim 14
Dependent Claim
18 words
The molecule of claim 1, wherein NA 1 or NA 2 is a naturally occurring nucleic acid sequence.
Claim 15
Dependent Claim
19 words
The molecule of claim 1, wherein a marker is attached to one of NA 1 or NA 2 .
Claim 16
Dependent Claim
10 words
The molecule of claim 14 immobilized on a solid support.
Claim 17
Dependent Claim
23 words
The molecule of claim 14, wherein the marker is a radioisotope, a radiolabelled molecule, a fluorescent molecule, biotin, an enzyme, or a ligand.
Claim 18
Dependent Claim
19 words
The molecule of claim 1, wherein NA 1 and NA 2 each comprise between 8 nucleotides and 10,000 nucleotides.
Claim 19
Dependent Claim
23 words
The molecule of claim 1, wherein n is greater than 1 and each [ NA 1 ---S---NA 2 ] unit is the same.
Claim 20
Dependent Claim
22 words
The molecule of claim 1, wherein n is greater than 1 and each ( NA 1 ---S---NA 2 ] unit is different.
Claim 21
Dependent Claim
34 words
A synthetic, non-naturally occurring molecule having the structure: wherein NA 1 and NA 2 are nucleic acid sequences; wherein ---S--- is a scissile linkage; and wherein n is an integer from 2 to 1000.
Claim 22
Dependent Claim
34 words
A synthetic, non-naturally occurring molecule having the structure: wherein NA 1 and NA 2 are nucleic acid sequences; wherein ---S--- is a scissile linkage; and wherein n is an integer from 2 to 1000.
Claim 23
Dependent Claim
19 words
The molecule of claim 20, wherein a marker is attached to one of NA 1 or NA 2 .
Claim 24
Dependent Claim
10 words
The molecule of claim 20 immobilized on a solid support.
Claim 25
Dependent Claim
63 words
A molecule having the structure: wherein NA 1 and NA 2 are nucleic acid sequences; wherein ---S--- is a scissile linkage; wherein n is an integer from 1 to 1,000; wherein the solid lines represent chemical bonds; wherein X is a solid support; wherein L is a chemical entity which links NA 1 to the solid support; and wherein M is a marker.
Claim 26
Dependent Claim
63 words
A molecule having the structure: wherein NA 1 and NA 2 are nucleic acid sequences; wherein ---S--- is a scissile linkage; wherein n is an integer from 1 to 1,000; wherein the solid lines represent chemical bonds; wherein X is a solid support; wherein L is a chemical entity which links NA 1 to the solid support; and wherein M is a marker.
Claim 27
Dependent Claim
18 words
The molecule of claim 23, wherein X is a silicaceous, cellulosic, or plastic material or controlled pore glass.
Claim 28
Dependent Claim
12 words
The molecule of claim 23, wherein the chemical bonds are covalent bonds.
Claim 29
Dependent Claim
14 words
The molecule of claim 23, wherein the chemical bonds are hydrogen or covalent bonds.
Claim 30
Dependent Claim
21 words
The molecule of claim 23, wherein M comprises a marked nucleic acid sequence complementary to a sequence within NA 2 .
Claim 31
Dependent Claim
19 words
The molecule of claim 23, wherein M is a radioisotope, a radiolabelled molecule, a fluorescent molecule, or a ligand.
Claim 32
Dependent Claim
12 words
The molecule of claim 23, wherein the dashed lines represent chemical bonds.
Claim 33
Dependent Claim
12 words
The molecule of claim 29, wherein the chemical bonds are covalent bonds.
Claim 34
Dependent Claim
12 words
The molecule of claim 29, wherein the chemical bonds are hydrogen bonds.
Claim 35
Dependent Claim
29 words
The molecule of claim 23, wherein the scissile linkage is selected from the group consisting of RNA sequences, DNA sequences, amino acid sequences, abasic nucleotide sequences or carbohydrate polymers.
Claim 36
Dependent Claim
19 words
The molecule of claim 23, wherein NA 1 and NA 2 each comprise between 8 nucleotides and 10,000 nucleotides.
Claim 37
Dependent Claim
24 words
The molecule of claim 23, wherein n is greater than 1 and each [ NA1 --- S --- NA2 ] unit is the same.
Claim 38
Dependent Claim
22 words
The molecule of claim 23, wherein n is greater than 1 and each [ NA 1 ---S---NA 2 ] unit is different.
Claim 39
Dependent Claim
51 words
The molecule of claim 33, wherein L comprises a nucleic acid which has a sequence substantially complementary to the sequence of NA,, wherein L is chemically bound to the solid support and wherein L links NA 1 to the solid support by means of hydrogen bonds between complementary nucleic acid sequences.
Claim 40
Dependent Claim
69 words
A molecule having the structure: wherein NA 1 and NA 2 are nucleic acid sequences; wherein ---S--- is a scissile linkage; wherein the solid lines represent chemical bonds; wherein n is an integer from 1 to 1,000; wherein Y is absent or is a chemical entity which confers a unique identifying characteristic; and wherein Z is absent or is a chemical entity which confers a different unique identifying characteristic.
Claim 41
Dependent Claim
69 words
A molecule having the structure: wherein NA 1 and NA 2 are nucleic acid sequences; wherein ---S--- is a scissile linkage; wherein the solid lines represent chemical bonds; wherein n is an integer from 1 to 1,000; wherein Y is absent or is a chemical entity which confers a unique identifying characteristic; and wherein Z is absent or is a chemical entity which confers a different unique identifying characteristic.
Claim 42
Dependent Claim
12 words
The molecule of claim 37, wherein the chemical bonds are covalent bonds.
Claim 43
Dependent Claim
12 words
The molecule of claim 37, wherein the chemical bonds are hydrogen bonds.
Claim 44
Dependent Claim
27 words
The molecule of claim 37, wherein when the identifying characteristic of Y is hydrophobicity or hydrophilicity and the identifying characteristic of Z is hydrophilicity or hydrophobicity, respectively.
Claim 45
Dependent Claim
18 words
The molecule of claim 37, wherein when one of Y or Z is absent, the other is hydrophobic.
Claim 46
Dependent Claim
12 words
The molecule of claim 37, wherein the dashed lines represent chemical bonds.
Claim 47
Dependent Claim
12 words
The molecule of claim 42, wherein the chemical bonds are covalent bonds.
Claim 48
Dependent Claim
12 words
The molecule of claim 42, wherein the chemical bonds are hydrogen bonds.
Claim 49
Dependent Claim
29 words
The molecule of claim 37, wherein the scissile linkage is selected from the group consisting of RNA sequences, DNA sequences, amino acid sequences, abasic nucleotide sequences or carbohydrate polymers.
Claim 50
Dependent Claim
19 words
The molecule of claim 37, wherein one of Y or Z is a marker and the other is hydrophobic.
Claim 51
Dependent Claim
19 words
The molecule of claim 37, wherein a marker is attached to one of NA 1 or NA 2 .
Claim 52
Dependent Claim
14 words
The molecule of claim 37, wherein NA 1 and NA 2 are DNA sequences.
Claim 53
Dependent Claim
14 words
The molecule of claim 37, wherein NA l and NA 2 are RNA sequences.
Claim 54
Dependent Claim
25 words
The molecule of claim 37, wherein NA 1 is either an RNA or DNA sequence and NA 2 is either an RNA or DNA sequence.
Claim 55
Dependent Claim
19 words
The molecule of claim 37, wherein NA 1 and NA 2 is not a naturally occurring nucleic acid sequence.
Claim 56
Dependent Claim
18 words
The molecule of claim 37, wherein NA 1 or NA 2 is a naturally occurring nucleic acid sequence.
Claim 57
Dependent Claim
14 words
A composition comprising a solid support and the molecule of claim 1 immobilized thereon.
Claim 58
Dependent Claim
92 words
A method of detecting in a sample the presence of a nucleic acid sequence of interest comprising: a) contacting the sample with a molecule of claim 14 which comprises a nucleic acid sequence substantially complementary to the nucleic acid sequence of interest, under hybridizing conditions; b) immobilizing the resulting molecule on a solid support; c) treating the immobilized molecule so as to cleave the scissile linkage; d) separately recovering the immobilized molecule; and e) detecting the presence of the marker on the immobilized molecule and thereby the nucleic acid sequence of interest.
Claim 59
Dependent Claim
83 words
A method of detecting in a sample the presence of a nucleic acid sequence of interest comprising: a) contacting the sample with a molecule of claim 23 which comprises a nucleic acid sequence substantially complementary to the nucleic acid sequence of interest, under hybridizing conditions; b) treating the immobilized molecule so as to cleave the scissile linkage; c) separately recovering the immobilized molecule; and d) detecting the presence of the marker on the immobilized molecule and thereby the nucleic acid sequence of interest.
Claim 60
Dependent Claim
133 words
A method of detecting in a sample a nucleic acid sequence of interest comprising: a) contacting under hybridizing conditions the sample with an aqueous solution comprising a molecule of claim 47 which comprises a nucleic acid sequence complementary to the nucleic acid sequence of interest and which includes a hydrophobic entity Y and a hydrophilic entity Z; b) mixing the resulting solution with a nonpolar solvent to form a biphasic solution in which the molecule is present at the phase interface of the biphasic solution, Y being oriented in one phase and Z being oriented in the other phase; c) treating the molecule so as to cleave the scissile linkage; d) recovering the molecule so treated; and e) detecting the presence of the marker and thereby detecting the nucleic acid sequence of interest.
Claim 61
Dependent Claim
132 words
A method of detecting in a sample a nucleic acid sequence of interest comprising: a) contacting under hybridizing conditions the sample with a molecule of claim 14 which comprises a nucleic acid sequence substantially complementary to the nucleic acid sequence of interest so as to form a hybridized product; b) contacting under hybridizing conditions the product of step (a) with a nucleic acid sequence which is complementary to the sequence of the molecule of claim 14, but has about 1-10 fewer bases and is capable of rendering the molecule undetectable when hybridized to the molecule, so that the nucleic acid sequence complementary to the molecule hybridizes with unhybridized molecule; and c) detecting the presence of the marker, thereby the presence of the hybridized product, and thereby the nucleic acid sequence of interest.
Claim 62
Dependent Claim
33 words
A diagnostic method for detecting the presence of a foreign pathogen in a sample which comprises detecting a nucleic acid sequence characteristic of the. pathogen using the method of any of claims 54-57.
Claim 63
Dependent Claim
100 words
A method for quantitatively determining in a sample the amount of a nucleic acid sequence of interest comprising: a) contacting the sample with a predetermined amount of a molecule of claim 14, which comprises a nucleic acid sequence substantially complementary to the nucleic acid sequence of interest, under hybridizing conditions; b) immobilizing the resulting molecule on a solid support; c) treating the immobilized molecule so as to cleave the scissile linkage; d) separately recovering the immobilized molecule; and e) quantitatively determining the amount of marker present on the immobilized molecule and thereby the amount of nucleic acid sequence of interest.
Claim 64
Dependent Claim
91 words
A method for quantitatively determining in a sample the amount of a nucleic acid sequence of interest comprising: a) contacting the sample with a predetermined amount of a molecule of claim 23, which comprises a nucleic acid sequence substantially complementary to the nucleic acid sequence of interest, under hybridizing conditions; b) treating the immobilized molecule so as to cleave the scissile linkage; c) separately recovering the immobilized molecule; and d) quantitatively determining the amount of marker present on the immobilized molecule and thereby the amount of nucleic acid sequence of interest.
Claim 65
Dependent Claim
136 words
A method for quantitatively determining in a sample the amount of a nucleic acid sequence of interest comprising: a) contacting under hybridizing conditions the sample with an aqueous solution comprising a molecule of claim 47 which comprises a nucleic acid sequence complementary to the nucleic acid sequence of interest and which includes a hydrophobic entity Y and a hydrophilic entity Z; b) mixing the resulting solution with a nonpolar solvent to form a biphasic solution in which the molecule is present at the phase interface of the biphasic solution, Y being oriented in one phase and Z being oriented in the other phase; c) treating the molecule so as to cleave the scissile linkage; d) recovering the molecule so treated; and e) quantitatively determining the amount of marker and thereby the nucleic acid sequence of interest.
Claim 66
Dependent Claim
31 words
A diagnostic method for measuring the amount of a foreign pathogen in a sample which comprises detecting a nucleic acid of the pathogen using the method of any of claims 59-61.
References Cited (Backward Citations)
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