Methods for labeling nucleotides, labeled nucleotides and useful intermediates
First Claim
1. A method for labeling a nucleotide comprising:
- providing a label;
providing a spacer means comprising a spacer reactive moiety at one end and an electron donating moiety at the other end of the spacer means , wherein the spacer reactive moiety is capable of coupling the spacer means to a label when the spacer reactive moiety is reacted with the label, wherein the spacer comprises a chain having at least four carbon atoms and at least one oxygen atom in the chain, and wherein the spacer comprises no more than twenty carbon atoms;
providing a nucleotide;
providing a linker having formula I,
wherein X represents an aliphatic diamine, and A and B represent the same or different linker reactive moieties capable of reacting with the electron donating group of the spacer means or with the nucleotide, thereby attaching the spacer means or the nucleotide to the linker;
reacting the spacer reactive moiety with the label, thereby coupling the spacer means to the label;
reacting the electron donating moiety of the spacer means with one of the linker reactive moieties, thereby attaching the spacer means to the linker; and
reacting the nucleotide with the other linker reactive moiety, thereby attaching the nucleotide to the linker.
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Abstract
The present invention provides improved methods for labeling nucleotides. The method of the invention for labeling nucleotides comprises the steps of: reacting a reactive moiety of a linker, which linker is a platinum compound having a stabilizing bridge and two reactive moieties, with an electron donating moiety of a spacer, which spacer comprises a chain having at least four atoms and at least one heteroatom in the chain, which spacer further comprises said electron donating moiety at one end of the chain and a reactive moiety at the other end of the chain; reacting the reactive moiety of said spacer with a label; reacting the other reactive moiety of said linker with a nucleotide. A major advantage of the invention is that all nucleotides can be labeled by the method of the invention, whereas until now the attachment of a label was mostly restricted to one or certain nucleotides.
1 Citation
28 Claims
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1. A method for labeling a nucleotide comprising:
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providing a label;
providing a spacer means comprising a spacer reactive moiety at one end and an electron donating moiety at the other end of the spacer means , wherein the spacer reactive moiety is capable of coupling the spacer means to a label when the spacer reactive moiety is reacted with the label, wherein the spacer comprises a chain having at least four carbon atoms and at least one oxygen atom in the chain, and wherein the spacer comprises no more than twenty carbon atoms;
providing a nucleotide;
providing a linker having formula I,
wherein X represents an aliphatic diamine, and A and B represent the same or different linker reactive moieties capable of reacting with the electron donating group of the spacer means or with the nucleotide, thereby attaching the spacer means or the nucleotide to the linker;
reacting the spacer reactive moiety with the label, thereby coupling the spacer means to the label;
reacting the electron donating moiety of the spacer means with one of the linker reactive moieties, thereby attaching the spacer means to the linker; and
reacting the nucleotide with the other linker reactive moiety, thereby attaching the nucleotide to the linker. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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Specification
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- Resources
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Current AssigneeKreatech Biotechnology BV (Danaher Corporation)
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Original AssigneeKreatech Biotechnology BV (Danaher Corporation)
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InventorsHeetebrij, Robert Jochem, Reedijk, Jan, Volkers, Herman Hendricus, Jelsma, Tinka, Houthoff, Jan Hendrick
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Primary Examiner(s)Sisson; Bradley L.
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Application NumberUS10/900,902Time in Patent Office4,038 DaysField of Search435/6, 435/91.1, 435/183, 435/287.2, 436/94, 436/800, 436/805, 536/23.1, 536/24.31, 536/24.33US Class Current435/6.1CPC Class CodesC07H 21/00 Compounds containing two or...C07H 23/00 Compounds containing boron,...Y10S 436/80 Fluorescent dyes, e.g. rhod...Y10T 436/143333 Saccharide [e.g., DNA, etc.]
