EP0122614A2
Expired / Lapsed
EP
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Kit for terminally chemically labeling DNA
Assignee: Enzo Biochem Inc
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Filing: 1984-04-12
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Granted: —
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Expires: 2004-04-12
Abstract
Abstract Processes and reagent kits employing terminal deoxynucleotide transferase are provided for terminally chemically labeling DNA probe molecules, such as the terminal or end labeling of DNA probes with biotinylated nucleotides, e.g. 5-alkylamino biotin-labeled dUTP.
Inventors
- 👤 Christine L. Brakel
- 👤 Jannis G. Stavrianopoulos
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 08:28
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
43 words
A reagent kit useful for the preparation of a terminally linked or labeled DNA molecule or probe comprising as reagents therein (1) terminal deoxyribonucleotide transferase TdT (2) Dnase and (3) a chemically labeled deoxyribonucleotide capable of acting as a substrate for said TdT.
Claim 2
Dependent Claim
43 words
A reagent kit useful for the preparation of a terminally linked or labeled DNA molecule or probe comprising as reagents therein (1) terminal deoxyribonucleotide transferase TdT (2) Dnase and (3) a chemically labeled deoxyribonucleotide capable of acting as a substrate for said TdT.
Claim 3
Dependent Claim
38 words
A reagent kit in accordance with Claim 1 wherein said chemically labeled deoxyribonucleotide is characterized such that when included in a DNA molecule said chemically labeled deoxyribonucleotide does not prevent said DNA molecule from forming double stranded DNA.
Claim 4
Dependent Claim
30 words
A reagent kit in accordance with Claim 1 wherein the chemical label moiety of said chemically labeled deoxyribonucleotide is capable of being detected by physical, physio-chemical, chemical or immunological techniques.
Claim 5
Dependent Claim
21 words
A reagent kit in accordance with Claim 1 wherein said chemically labeled deoxyribonucleotide is a biotinylated deoxyribonucleotide, especially 2'-deoxyuridine triphosphate 5-allylamine-biotin.
Claim 6
Dependent Claim
20 words
5 .. A reagent kit in accordance with Claim 1 comprising additionally an aqueous COCl 2 and/or MgC1 2 solution.
Claim 7
Dependent Claim
29 words
6 . A reagent kit in accordance with Claim 1 comprising additionally as Dnase dilution buffer a solution comprising 10 mM Tris HC1 of a pH of about 7.5.
Claim 8
Dependent Claim
25 words
7 . A reagent kit in accordance with Claim 1 comprising additionally for stabilizing the Dnase a solution containing about 1 mg/ml bovine serum albumin.
Claim 9
Dependent Claim
78 words
A method of preparing terminally linked or labeled DNA employing the reagent kit of Claim 1 which comprises bringing DNA to be terminally linked or labeled, after having been treated with said Dnase, into contact with said chemically labeled deoxyribonucleotide in the presence of said TdT to effect terminal attachment of said chemically labeled deoxyribonucleotide to the 3' OH position presented by said DNA or to any chemically labeled deoxyribonucleotide previously attached to said DNA via said TdT.
Claim 10
Dependent Claim
52 words
A reagent kit useful for the preparation of a terminally linked or labeled DNA molecule or probe comprising as reagents therein (1) terminal deoxyribonucleotide transferase TdT (2) an enzyme capable of producing or exposing 3'-OH termini of DNA and (3) a chemically labeled deoxyribonucleotide capable of acting as a substrate for TdT.
Claim 11
Dependent Claim
52 words
A reagent kit useful for the preparation of a terminally linked or labeled DNA molecule or probe comprising as reagents therein (1) terminal deoxyribonucleotide transferase TdT (2) an enzyme capable of producing or exposing 3'-OH termini of DNA and (3) a chemically labeled deoxyribonucleotide capable of acting as a substrate for TdT.
Claim 12
Dependent Claim
79 words
A method of preparing terminally linked or labeled DNA employing the reagent kit of Claim 9, which comprises bringing DNA to be terminally linked or labeled, after having been treated with said enzyme, into contact with said chemically labeled deoxyribonucleotide in the presence of said TdT to effect terminal attachment of said chemicaly labeled deoxyribonucleotide to the 3'-OH position or termini presented by said DNA or to any chemically labeled deoxyribonucleotide previously attached to said DNA via said TdT.
Claim 13
Dependent Claim
28 words
11 . A terminally labeled DNA molecule or probe wherein said DNA molecule or probe is terminally linked or labeled with at least one non-radioactive chemically labeled deoxyribonucleotide.
Claim 14
Dependent Claim
28 words
11 . A terminally labeled DNA molecule or probe wherein said DNA molecule or probe is terminally linked or labeled with at least one non-radioactive chemically labeled deoxyribonucleotide.
Claim 15
Dependent Claim
31 words
12 A terminally labeled DNA molecule or probe in accordance with Claim 11 wherein said non-radioactive chemically labeled deoxyribonucleotide is capable of acting as a substrate for terminal deoxyribonucleotide transferase TdT.
Claim 16
Dependent Claim
35 words
A terminally labeled DNA molecule or probe in accordance with claim 11 wherein said non-radioactive chemically labeled desoxyribonucleotide is a desoxyribonucleotide as used in the reagent kit according to anyone of claims 1 to 4.
Claim 17
Dependent Claim
21 words
A terminally labeled DNA molecule or probe in accordance with Claim 11. wherein said chemically labeled deoxyribonucleotide is a glycosylated deoxyribonucleotide.
Claim 18
Dependent Claim
34 words
A terminally labeled DNA molecule or probe wherein said DNA-molecule or probe is terminally linked or labeled with a single stranded polydeoxyribonucleotide, said single stranded polydeoxyribonucleotide containing one or more non-radioactive chemically labeled deoxyribonucleotides.
Claim 19
Dependent Claim
34 words
A terminally labeled DNA molecule or probe wherein said DNA-molecule or probe is terminally linked or labeled with a single stranded polydeoxyribonucleotide, said single stranded polydeoxyribonucleotide containing one or more non-radioactive chemically labeled deoxyribonucleotides.
Claim 20
Dependent Claim
21 words
A chemically labeled deoxyribonucleotide in accordance with Claim 15 wherein said chemically labeled deoxyribonucleotide is a biotinylated or a glycosylated desoxyribonucleotide.
Claim 21
Dependent Claim
18 words
A chemically labeled deoxyribonucleotide in accordance with Claim 16 wherein said chemically labeled deoxyribonucleotide is 2'-deoxyuridine triphosphate 5-allylamine-biotin.
Claim 22
Dependent Claim
39 words
A chemically labeled deoxyribonucleotide in accordance with Claim 15 wherein said chemically labeled deoxyribonucleotide comprises from about 1 % up to 100% , preferably from about 2% to about 40% of the nucleotides making up said single stranded polydeoxyribonucleotide.
References Cited (Backward Citations)
No citation records indexed.
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