DE164054T1 - Verfahren zum nachweis von polymorphischen restriktionsstellen und nukleinsauren sequenzen. - Google Patents
Verfahren zum nachweis von polymorphischen restriktionsstellen und nukleinsauren sequenzen.Info
- Publication number
- DE164054T1 DE164054T1 DE198585106525T DE85106525T DE164054T1 DE 164054 T1 DE164054 T1 DE 164054T1 DE 198585106525 T DE198585106525 T DE 198585106525T DE 85106525 T DE85106525 T DE 85106525T DE 164054 T1 DE164054 T1 DE 164054T1
- Authority
- DE
- Germany
- Prior art keywords
- restriction site
- sample
- restriction
- nucleic acid
- acid sequence
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
- C12Q1/683—Hybridisation assays for detection of mutation or polymorphism involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6832—Enhancement of hybridisation reaction
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/803—Physical recovery methods, e.g. chromatography, grinding
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/81—Packaged device or kit
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/807—Apparatus included in process claim, e.g. physical support structures
- Y10S436/808—Automated or kit
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/811—Test for named disease, body condition or organ function
Claims (8)
1. Verfahren zum Nachweis der Anwesenheit oder Abwesenheit mindestens einer bestimmten Restriktionsstelle in einer
bestimmten Nukleinsäure-Sequenz, gekennzeichnet durch die Stufen:
(a) Hybridisierung der Nukleinsäure-Sequenz mit einer Oligonukleotid-Probe für jede nachzuweisende Restriktionsstelle, wobei die Probe zu einem Bereich der Nukleinsäure-Sequenz
komplementär ist, der die entsprechende Restrik-
tionsstelle der Probe überspannt und wobei die Probe an
dem der jeweiligen Restriktionsstelle näheren Ende markiert ist;
(b) Verdauen der hybridisierten Nukleinsäure-Sequenz mit einer Restriktions-Endonuklease für jede Probe, wel-
ehe zur Spaltung der jeweiligen Probe an der nachzuweisenden
Restriktionsstelle befähigt ist, wobei markierte und unmarkierte oligomere Fragmente erhalten werden, bei
denen die Markierung die gleiche ist wie diejenige auf der Probe;
(c) Abtrennen aller markierten, gespalteten oligomeren Fragmente von markierten nichtgespalteten Oligomeren;
und
(d) Nachweis der Gegenwart oder Abwesenheit von markierten oligomeren Fragmenten.
35
L J
Γ *■·'■·· ■-•-»2--·..·'..· 0164052
2. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß
es ein Verfahren zum Nachweis der Gegenwart einer Restriktionsstelle ist, deren Bezug zu einer Infektionskrankheit
bekannt ist.
3. Verfahren zum Nachweis der Gegenwart oder Abwesenheit von einem oder beiden Allelen einer polymorphen Restriktionsstelle in einer bestimmten Nukleinsäure-Sequenz, gekennzeichnet
durch die Stufen:
(a) Hybridisierung der Nukleinsäure-Sequenz mit einer Oligonukleotid-Probe, welche komplementär ist zu einem
Bereich in der Nukleinsäure-Sequenz, der eine nur in einem Alle! vorhandene erste Restriktionsstelle und eine beiden
Allelen gemeinsame zweite Restriktionsstelle überspannt, und wobei die Probe an einem Ende markiert ist, das sich
näher bei einer der genannten Restrictionsstellen befindet
;
(b) Verdauen der hybridisierten Nukleinsäure-Sequenz mit einer ersten Restriktionsendonuklease, welche zur Spaltung
der Probe nur an der ersten Restriktionsstelle befähigt ist, wobei markierte und unmarkierte oligomere
Fragmente erhalten werden, in denen die Markierung die gleiche wie diejenige an der Probe ist;
(c) Verdauen des Verdaus von Stufe (b) mit einer zweiten
Restriktionsendonuklease, welche zur Spaltung der Probe nur an der zweiten Restriktionsstelle befähigt ist, wobei
markierte und unmarkierte oligomere Fragmente erhalten werden, in denen die Markierung die gleiche wie diejenige
der Probe ist;
(d) Abtrennung aller markierten, gespalteten oligomeren
Fragmente von nichtgespalteten Oligomeren; und (e) Nachweis der Gegenwart oder Abwesenheit von markierten
Fragmenten mit bestimmten Basenlängen, die mit der Spaltung durch die erste oder zweite Restriktionsendonuklease
übereinstimmen.
4. Verfahren nach Anspruch 5? dadurch gekennzeichnet, daß
die Krankheit Sichelzellanämie ist, die erste Sestriktionsstelle die in normalem ß-Globin vorhandene, in Sichelzellen-fi-Globin
jedoch abwesende Ddel-Schnittstelle
ist, und die zweite Restriktionsstelle, die sowohl in normalem als auch in Sichelzellen-ß-Globin vorhandene
Hinf!-Schnittstelle ist.
5. "Verfahren nach einem der Ansprüche 1 bis 4, weiter da-
durch gekennzeichnet, daß nach der Stufe (a) und vor der Stufe (b) das Verfahren gekennzeichnet ist durch die
Stufen:
(a1) Versetzen des Hybridisierungsgemisches von Stufe (a)
mit einem unmarkierten blockierenden Oligomeren, das zu allen Proben mit Ausnahme von mindestens einem nicht richtig
zusammenpassenden Basenpaar an jeder nachzuweisenden Restriktionsstelle komplementär ist; und
(a") Hybridisieren des Hybridisierungsgemisches in Gegenwart des blockierenden Oligomeren.
6. Verfahren nach einem der Ansprüche 1 bis 5, dadurch gekennzeichnet,
daß die Restriktionsstelle polymorph ist und die Hybridisierungsstufe in Gegenwart eines positiven
Vergleichs-Oligomeren durchgeführt wird, das an einen Bereich der Nukleinsäure-Sequenz bindet, welcher eine nichtpolymorphe
Restriktionsstelle für die Restriktionsendonuklease enthält.
7· Verfahren nach einem der Ansprüche 1 bis 6, dadurch gekennzeichnet,
daß die Nukleinsäure-Sequenz eine DNA-Sequenz ist und die Probe ein Oligodeoxyribonukleotid ist.
8. Nachweissatz zum raschen Nachweis der Gegenwart oder Abwesenheit
mindestens einer bestimmten Restriktionsstelle in einer bestimmten Nukleinsäure-Sequenz, gekennzeichnet
durch:
L J
- ·ι+- 01
(a) eine Cligonukleotid-Probe für ,jede nachzuweisende Restriktionsstelle,
wobei die Probe zu einem Bereich der Nukleinsäure-Sequenz komplementär ist, der die betreffende
Restrik~ionsstelle der Probe überspannt, und wobei die
Probe an dem der betreffenden Restriktionsstelle näheren
Ende markiert ist; und
(b) eine Restriktionsendonuklease für jede Probe, die zur
Spaltung der Probe an der Restriktionsstelie befähigt ist.
9· Nachweissatz nach Anspruch 8, weiter gekennzeichnet durch
ein nichtmarkiertes blockierendes Oligomeres, das zu allen Proben mit Ausnahme von mindestens einem nichtpassenden
Basenpaar an der Restriktionsstelle komplementär ist.
1C. Nachweissatz zum Nachweis der Gegenwart oder Abwesenheit von einem oder beiden Allelen einer poiymorphen Restriktionsstelle
in einer bestimmten DNA-Sequenz, gekennzeichnet durch:
(a) eine Oligodeoxyribonukleotid-Probe, welche zu einem Bereich in der DNA-Sequenz komplementär ist, der eine erste, nur in einem Allel vorhandene Restriktionsstelle und eine zweite, beiden Allelen gemeinsame Restriktionsstellle überspannt und wobei die Probe an dem zu einer der Restriktionsstellen näheren Ende markiert ist; ("b) eine erste Restriktionsendonuklease, die zur Spaltung der Probe nur an der ersten Restriktionsstelle befähigt ist; und
(a) eine Oligodeoxyribonukleotid-Probe, welche zu einem Bereich in der DNA-Sequenz komplementär ist, der eine erste, nur in einem Allel vorhandene Restriktionsstelle und eine zweite, beiden Allelen gemeinsame Restriktionsstellle überspannt und wobei die Probe an dem zu einer der Restriktionsstellen näheren Ende markiert ist; ("b) eine erste Restriktionsendonuklease, die zur Spaltung der Probe nur an der ersten Restriktionsstelle befähigt ist; und
(c) eine zweite Restriktionsendonuklease, die zur Spaltung der Probe nur an der zweiten Restriktionsstelle
befähigt ist.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US61495784A | 1984-05-29 | 1984-05-29 | |
US06/716,982 US4683194A (en) | 1984-05-29 | 1985-03-28 | Method for detection of polymorphic restriction sites and nucleic acid sequences |
Publications (1)
Publication Number | Publication Date |
---|---|
DE164054T1 true DE164054T1 (de) | 1986-04-30 |
Family
ID=27087371
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DE198585106525T Pending DE164054T1 (de) | 1984-05-29 | 1985-05-28 | Verfahren zum nachweis von polymorphischen restriktionsstellen und nukleinsauren sequenzen. |
DE8585106525T Expired DE3572693D1 (en) | 1984-05-29 | 1985-05-28 | Method for detection of polymorphic restriction sites and nucleic acid sequences |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DE8585106525T Expired DE3572693D1 (en) | 1984-05-29 | 1985-05-28 | Method for detection of polymorphic restriction sites and nucleic acid sequences |
Country Status (9)
Country | Link |
---|---|
US (1) | US4683194A (de) |
EP (1) | EP0164054B1 (de) |
JP (1) | JPS6119500A (de) |
AU (1) | AU587189B2 (de) |
CA (1) | CA1242627A (de) |
DE (2) | DE164054T1 (de) |
DK (1) | DK240285A (de) |
ES (1) | ES8608584A1 (de) |
IL (1) | IL75327A (de) |
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US4477577A (en) * | 1980-03-17 | 1984-10-16 | University Of Southern California | Non-contaminating direct serum assay of steroid hormones |
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US4745060A (en) * | 1984-12-28 | 1988-05-17 | Board Of Regents, The University Of Texas System | Methods and compositions for the detection of Familial Hypercholesterolemia |
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1985
- 1985-03-28 US US06/716,982 patent/US4683194A/en not_active Expired - Fee Related
- 1985-04-29 CA CA000480314A patent/CA1242627A/en not_active Expired
- 1985-05-28 DE DE198585106525T patent/DE164054T1/de active Pending
- 1985-05-28 EP EP85106525A patent/EP0164054B1/de not_active Expired
- 1985-05-28 IL IL75327A patent/IL75327A/xx unknown
- 1985-05-28 DE DE8585106525T patent/DE3572693D1/de not_active Expired
- 1985-05-28 ES ES543547A patent/ES8608584A1/es not_active Expired
- 1985-05-29 AU AU43083/85A patent/AU587189B2/en not_active Ceased
- 1985-05-29 JP JP60114408A patent/JPS6119500A/ja active Pending
- 1985-05-29 DK DK240285A patent/DK240285A/da not_active Application Discontinuation
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ES8608584A1 (es) | 1986-06-16 |
AU4308385A (en) | 1985-12-05 |
DK240285A (da) | 1985-11-30 |
EP0164054B1 (de) | 1989-08-30 |
ES543547A0 (es) | 1986-06-16 |
US4683194A (en) | 1987-07-28 |
IL75327A (en) | 1990-02-09 |
EP0164054A1 (de) | 1985-12-11 |
IL75327A0 (en) | 1985-09-29 |
DK240285D0 (da) | 1985-05-29 |
CA1242627A (en) | 1988-10-04 |
JPS6119500A (ja) | 1986-01-28 |
DE3572693D1 (en) | 1989-10-05 |
AU587189B2 (en) | 1989-08-10 |
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