EP0238329A2
Expired / Lapsed
EP
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Improvements in genetic probes
Assignee: Syngenta Ltd
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Filing: 1987-03-18
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Granted: —
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Expires: 2007-03-18
Abstract
Abstract Test samples of genomic DNA may be characterised by the use of polynucleotide probes each of which is specific for an informative genetic locus. Such probes may be prepared by the use of probes which are capable of differentiating DNA by reference to more than one polymorphic minisatellite region or hypervariable locus. The polynucleotides and probes of the invention are of use for genetic identification purposes, paternity and maternity testing and particularly in forensic medicine.
Inventors
- 👤 Alec John Jeffreys
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-06 02:55
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6827; C12Q1/6876; C12Q2600/00; C12Q2600/156; C12Q2600/172
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
144 words
A method of preparing a polynucleotide capable of hybridising to a DNA fragment which contains a minisatellite which is specific as to a particular region or locus in the genome, said region or locus having an allelic variation (as herein defined) of at least 3(100) or a degree of polymorphism of at least 3; which method comprises cloning a DNA fragment identified by hybridisation of fragments of genomic DNA with a polynucleotide probe which is capable of differentiating DNA by reference to more than one polymorphic minisatellite region or hypervariable locus; and preparing therefrom a polynucleotide capable of hybridising to a DNA fragment which contains a minisatellite which is specific as to a particular region or locus in the genome, said region or locus having an allelic variation (as herein defined) of at least 3(100) or a degree of polymorphism of at least 3.
Claim 2
Independent Claim
144 words
A method of preparing a polynucleotide capable of hybridising to a DNA fragment which contains a minisatellite which is specific as to a particular region or locus in the genome, said region or locus having an allelic variation (as herein defined) of at least 3(100) or a degree of polymorphism of at least 3; which method comprises cloning a DNA fragment identified by hybridisation of fragments of genomic DNA with a polynucleotide probe which is capable of differentiating DNA by reference to more than one polymorphic minisatellite region or hypervariable locus; and preparing therefrom a polynucleotide capable of hybridising to a DNA fragment which contains a minisatellite which is specific as to a particular region or locus in the genome, said region or locus having an allelic variation (as herein defined) of at least 3(100) or a degree of polymorphism of at least 3.
Claim 3
Dependent Claim
146 words
A polynucleotide, in isolated or cloned form, which comprises a nucleotide sequence which is specific as to a single minisatellite region or hypervariable locus and which is homologous therewith to a degree enabling hybridisation of the nucleotide sequence to a corresponding DNA fragment obtained by fragmenting sample genomic DNA with a restriction endonuclease, the said DNA fragment containing a minisatellite from said minisatellite region or hypervariable locus, wherein:- 1) said region or locus has an allelic variation (as herein defined) of at least 3(100) or a degree of polymorphism of at least 3; and 2) said region or locus is detectable by a polynucleotide probe comprising at least three tandem repeats of the sequence (including complementary sequences):- (A)AGGGCTGGAGG 1 (wherein T is T or U and (A) may be present or absent) or the sequence (including complementary sequences):- AGAGGTGGGCAGGTGG 2 (wherein T is T or U).
Claim 4
Dependent Claim
146 words
A polynucleotide, in isolated or cloned form, which comprises a nucleotide sequence which is specific as to a single minisatellite region or hypervariable locus and which is homologous therewith to a degree enabling hybridisation of the nucleotide sequence to a corresponding DNA fragment obtained by fragmenting sample genomic DNA with a restriction endonuclease, the said DNA fragment containing a minisatellite from said minisatellite region or hypervariable locus, wherein:- 1) said region or locus has an allelic variation (as herein defined) of at least 3(100) or a degree of polymorphism of at least 3; and 2) said region or locus is detectable by a polynucleotide probe comprising at least three tandem repeats of the sequence (including complementary sequences):- (A)AGGGCTGGAGG 1 (wherein T is T or U and (A) may be present or absent) or the sequence (including complementary sequences):- AGAGGTGGGCAGGTGG 2 (wherein T is T or U).
Claim 5
Dependent Claim
38 words
A process of preparing a polynucleotide first prepared by a method as defined in claim 1 or of preparing a polynucleotide as defined in claim 2, which process comprises direct synthesis or the microbiological reproduction of cloned material.
Claim 6
Dependent Claim
31 words
A polynucleotide probe which comprises a polynucleotide as defined in claim 2 or as prepared by a method as defined in claim 1, said polynucleotide having a labelled or marker component.
Claim 7
Dependent Claim
35 words
A method of preparing a polynucleotide probe as defined in claim 4 which comprises labelling or marking a polynucleotide as claimed of claims 2 or as prepared by a method as defined in claim 1.
Claim 8
Dependent Claim
170 words
A method of preparing a polynucleotide capable of hybridising to a DNA fragment which contains a minisatellite which is specific as to a particular region or locus in the genome, said region or locus having an allelic variation (as herein defined) of at least 3(100) or a degree of polymorphism of at least 3. which method comprises cloning a DNA fragment identified by hybridisation of fragments of genomic DNA with a polynucleotide probe as claimed in claim 4 or a polynucleotide probe prepared by the method of claim 5, and preparing therefrom a polynucleotide capable of hybridising to a DNA fragment which contains a minisatellite which is specific as to a particular region or locus on the genome, said region or locus having an allelic variation (as herein defined) of at least 3(100) or a degree of polymorphism of at least 3; the method being effected under hybridisation conditions effective to render the probe capable of differentiating DNA by reference to more than one polymorphic minisatellite region or hypervariable locus.
Claim 9
Dependent Claim
119 words
A polynucleotide, in isolated or cloned form, which comprises a nucleotide sequence which is specific as to a single minisatellite region or hypervariable locus and which is homologous therewith to a degree enabling hybridisation of the nucleotide sequence to a corresponding DNA fragment obtained by fragmenting sample genomic DNA with a restriction endonuclease, the said DNA fragment containing a minisatellite from said minisatellite region or hypervariable locus, wherein:- 1) said region or locus has an allelic variation (as herein defined) of at least 3(100) or a degree of polymorphism of at least 3; and 2) said region or locus is detectable by a polynucleotide probe as claim 4 or a polynucleotide probe prepared by the method of claim 5.
Claim 10
Independent Claim
171 words
A method of characterising a test sample of genomic DNA by reference to one or more controls which method comprises fragmenting sample DNA with one or more restriction enzymes which do not cleave to any relevant extent a sequence corresponding to a tandem repeat, probing the DNA fragments with a polynucleotide or polynucleotide probe comprising a nucleotide sequence which is specific as to a single minisatellite region or hypervariable locus and which is homologous therewith to a degree enabling hybridisation of the nucleotide sequence to a corresponding DNA fragment containing a minisatellite from said single minisatellite region or hypervariable locus, detecting hybridised fragments of DNA, and comprising the hybridised fragments with a said control or controls wherein:- 1) said minisatellite region or hypervariable locus has an allelic variation of at least 3(100) or a degree of polymorphism of at least 3; and 2) said region or locus is detectable by a polynucleotide probe which is capable of differentiating DNA by reference to more than one polymorphic minisatellite region or hypervariable locus.
Claim 11
Independent Claim
171 words
A method of characterising a test sample of genomic DNA by reference to one or more controls which method comprises fragmenting sample DNA with one or more restriction enzymes which do not cleave to any relevant extent a sequence corresponding to a tandem repeat, probing the DNA fragments with a polynucleotide or polynucleotide probe comprising a nucleotide sequence which is specific as to a single minisatellite region or hypervariable locus and which is homologous therewith to a degree enabling hybridisation of the nucleotide sequence to a corresponding DNA fragment containing a minisatellite from said single minisatellite region or hypervariable locus, detecting hybridised fragments of DNA, and comprising the hybridised fragments with a said control or controls wherein:- 1) said minisatellite region or hypervariable locus has an allelic variation of at least 3(100) or a degree of polymorphism of at least 3; and 2) said region or locus is detectable by a polynucleotide probe which is capable of differentiating DNA by reference to more than one polymorphic minisatellite region or hypervariable locus.
Claim 12
Dependent Claim
45 words
A method according to claim 8 of establishing the identity or otherwise of the test sample donor with one or more control sample donors, in which the control sample(s) are similarly fragmented and a comparison is made of the hybridised fragments from the respective samples.
Claim 13
Dependent Claim
55 words
A polynucleotide probe which comprises any one sequence selected from AGGAATAGAAAGGCGGGYGGTGTGGGCAGGGAGRGGC 3 GTGGAYAGG 4 TGGGAGGTGGRYAGTGTCTG 5 GAATGGAGCAGGYGRCCAGGGGTGACTCA 6 GGGCTGGGGAGATGGTGGAGGAGGTGTTGG 7 AGGCTGGGGAGATGGTGGAGGAAGAGTAC 8 TGTGTGTAATGGGTATAGGGAGGGCCCCGGGAAGGGGGTGTGGYX 9 (wherein Y is C, T or U, X is G or C, R is A or G and T is T or U) or tandem repeat(s) thereof or sequence(s) complementary thereto.
Claim 14
Dependent Claim
55 words
A polynucleotide probe which comprises any one sequence selected from AGGAATAGAAAGGCGGGYGGTGTGGGCAGGGAGRGGC 3 GTGGAYAGG 4 TGGGAGGTGGRYAGTGTCTG 5 GAATGGAGCAGGYGRCCAGGGGTGACTCA 6 GGGCTGGGGAGATGGTGGAGGAGGTGTTGG 7 AGGCTGGGGAGATGGTGGAGGAAGAGTAC 8 TGTGTGTAATGGGTATAGGGAGGGCCCCGGGAAGGGGGTGTGGYX 9 (wherein Y is C, T or U, X is G or C, R is A or G and T is T or U) or tandem repeat(s) thereof or sequence(s) complementary thereto.
Claim 15
Dependent Claim
20 words
A polynucleotide probe which comprises the sequences of formula 3 (as defined in claim 10) or a tandem repeat thereof.
Claim 16
Dependent Claim
81 words
A method of characterising a test sample of genomic DNA by reference to one or more controls which method comprises fragmenting sample DNA with one or more restriction enzymes which do not cleave to any relevant extent a sequence corresponding to a tandem repeat, probing the DNA fragments with a polynucleotide probe as defined in claim 10 or claim 11 detecting fragments of DNA which have hybridised with the probe and comparing the hybridised fragments with a said control or controls.
References Cited (Backward Citations)
GB2166445A
Cited
EP0201184A2
Cited
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