Patent EP0266787A2 successfully refreshed from Google Patents.
EP0266787A2
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EP
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Process for the detection of restriction fragment length polymorphisms in eukaryotic genomes
Assignee: Max-Planck-Gesellschaft zur Förderung der Wissenschaften eV
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Filing: 1987-11-06
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Granted: —
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Expires: 2007-11-06
Abstract
Abstract The invention is directed to a process for the analysis of eukaryotic DNA wherein DNA of an individual is isolated, the DNA is subjected to restriction endonuclease digest, the cleaved DNA is separated by gel electrophoresis, the gel is dried and hybridized with a probe, and the pattern of the DNA is evaluated. The process is characterized in that the probe used is an oligonucleotide which specifically hybridizes to eukaryotic simple tandem repeats having a periodicity of 2 to 4 bases. The process of the invention may be employed for the in vitro diagnosis of genetic defects such as hereditary diseases or for the determination of the genetic interrelationships.
Inventors
- 👤 Jörg T. Dr. Epplen
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 06:03
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12Q1/6813; C12Q1/6827; C12Q1/683; C12Q2600/00; C12Q2600/156
Claims Analysis
Total Claims: 35
Claim 1
Independent Claim
73 words
A process for the analysis of eukaryotic DNA wherein DNA of an individual is isolated, the DNA is subjected to restriction endonuclease digest, the cleaved DNA is separated by gel electrophoresis, the gel is dried and hybridized with a probe, and the pattern of the DNA is evaluated, characterized in that the probe used is an oligonucleotide which specifically hybridizes to eukaryotic simple tandem repeats having a periodicity of 2 to 4 bases.
Claim 2
Dependent Claim
73 words
A process for the analysis of eukaryotic DNA wherein DNA of an individual is isolated, the DNA is subjected to restriction endonuclease digest, the cleaved DNA is separated by gel electrophoresis, the gel is dried and hybridized with a probe, and the pattern of the DNA is evaluated, characterized in that the probe used is an oligonucleotide which specifically hybridizes to eukaryotic simple tandem repeats having a periodicity of 2 to 4 bases.
Claim 3
Dependent Claim
17 words
The process according to claim 1, characterized in that the DNA used is human or animal DNA.
Claim 4
Dependent Claim
28 words
The process according to any one of claims 1 and 2, characterized in that the restrictions enzyme used is Alu I, Hae III, Hinf I, or Mbo I.
Claim 5
Dependent Claim
24 words
The process according to claim 1, characterized in that the probe used is an oligonucleotide which specifically hybridizes to eukaryotic simple quadruplet repeats (sqr).
Claim 6
Dependent Claim
32 words
The process according to claim 4. characterized in that the simple quadruplet repeats (sar) recognized by the oligonucleotide probe in the eukaryotic genome are GA C A sequences or their complement sequences
Claim 7
Dependent Claim
19 words
The process according to claim 5, characterized in that the oligonucleotide has a length of 16 to 23 nucleotides.
Claim 8
Dependent Claim
20 words
The process according to claim 6, characterized in that the oligonucleotide is (GATA)aGA or its complement sequence TC(TATC) 4 .
Claim 9
Dependent Claim
14 words
The process according to claim 6, characterized in that the nucleotide is a 20-mer.
Claim 10
Dependent Claim
21 words
The process according to claim 6, characterized in that the oligonucleotide is (GATA) 5 or its complement sequence (TATC) 5 .
Claim 11
Dependent Claim
26 words
The process according to claim 6. characterized in that the oligonucleotide is (GATA) 2 GACA (GATA) 2 or its complement sequence (TATC) 2 TGTC(TATC) 2 .
Claim 12
Dependent Claim
27 words
The process according to claim 6. characterized in that the oligonucleotide is (GATA) 4 . (GACA) 4 . or their complement sequences (TATC)4 or (TGTC) 4 .
Claim 13
Dependent Claim
21 words
The process according to claim 6, characterized in that the oligonucleotide is (GGAT) 2 or its complement sequence (ATCC) 2 .
Claim 14
Dependent Claim
34 words
The process according to claim 6, characterized in that the oligonucleotide has one of the sequences as mentioned in claims 6, 8. 9, or 10 but beginning with any one of the other nucleotides.
Claim 15
Dependent Claim
52 words
The process according to any one of claims 1 to 11 characterized in that several probes of different length within the range of 16 to 23 nucleotides are used in consecutive hybridization and evaluation operations, the hybridization with the probes being carried out at the specific hybridization temperature of the individual probe.
Claim 16
Dependent Claim
15 words
Process according to claim 14, characterized in that more than one digestion is carried out.
Claim 17
Dependent Claim
32 words
Process according to claims 14 and 15, characterized in that one of the probes used is (GATA)aGA having a hybridization temperature of about 41 °C or its complement sequence TC(TATC) 4 .
Claim 18
Dependent Claim
32 words
Process according to claims 14 and 15, characterized in that one of the probes used is (GATA) 5 having a hybridization temperature of about 45°C or its complement sequence (TATC) 5 .
Claim 19
Dependent Claim
36 words
Process according to claims 14 and 15, characterized in that one of the probes used is (GATA)- 2 GACA(GATA) 2 having a hybridization temperature of about 47°C or its complement sequence (TATC)- 2 TGTC(TATC) 2 .
Claim 20
Dependent Claim
41 words
Process according to claims 14 and 15, characterized in that one of the probes used is (GATA), or (GACA) 4 having a hybridization temperature of about 35°C and about 43°C, respectively, or its complement sequence (TATC) 4 or (TGTC) 4 .
Claim 21
Dependent Claim
23 words
The process according to claim 1, characterized in that the probe used is an oligonucleotide which specifically hybridizes to eukaryotic simple triplet repeats.
Claim 22
Dependent Claim
30 words
The process according to claim 20, characterized in that the simple triplet repeats recognized by the oligonucleotide probe in the eukaryotic genome are TCC sequences or their complement sequences GGA.
Claim 23
Dependent Claim
19 words
The process according to claim 21, characterized in that the oligonucleotide has a length of 15 to 24 nucleotides.
Claim 24
Dependent Claim
21 words
The process according to claim 22, characterized in that the oligonucleotide is (TCC) 5 or its complement sequence (GGA) 5 .
Claim 25
Dependent Claim
28 words
The process according to claim 22, characterized in that the oligonucleotide has the sequence as mentioned in claim 23 but beginning with any one of the other nucleotides.
Claim 26
Dependent Claim
23 words
The process according to claim 1, characterized in that the probe used is an oligonucleotide which specifically hybridizes to eukaryotic simple doublet repeats.
Claim 27
Dependent Claim
30 words
The process according to claim 25, characterized in that the simple duplet repeats recognised by the oligonucleotide probe in the eukaryotic genome are GT sequences or their complement sequences AC.
Claim 28
Dependent Claim
19 words
The process according to claim 26, characterized in that the oligonucleotide has a length of 16 to 24 nucleotides.
Claim 29
Dependent Claim
20 words
The process according to claim 27, characterized in that the oligonucleotide is (GT): or its complement sequence (AC) 1 .
Claim 30
Dependent Claim
20 words
The process according to claim 27, characterized in that the oligonucleotide is (CT): or its complement sequence (AG) 8 .
Claim 31
Dependent Claim
27 words
The process according to claim 27, characterized in that the oligonucleotide has the sequence as mentioned in claims 28 or 29 but beginning with the other nucleotide.
Claim 32
Dependent Claim
54 words
The process according to claim 1. characterized in that the consecutive hybridization and evaluation operation comprises hybridizing the cleaved DNA with the radioactively labeled probe, removing the unhybridized part of the probe, exposing an X-ray film to the hybridized product, removing the hybridized product from the gel and repeating the operation with another probe.
Claim 33
Dependent Claim
37 words
The process according to claim 1, characterized in that the hybridization and evaluation operation comprises transferring the cleaved DNA onto a solid support hybridizing the DNA with the biotinylated probe and developing the hybridized product with strepdavidin.
Claim 34
Dependent Claim
27 words
A process for the in vitro diagnosis of genetic defects such as hereditary diseases comprising a DNA analysis according to any one of claims 1 to 30.
Claim 35
Dependent Claim
22 words
A process for the determination of the genetical interrelationships comprising a DNA analysis according to any one of claims 1 to 30.
References Cited (Backward Citations)
No citation records indexed.
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