Processes using nucleoside triphosphates with stable aminoxy groups
AssigneeFirebird Biomolecular Sciences
InventorsNicole A. Leal, Steven A Benner
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Abstract
This invention claims processes that append a single nucleotide having a 3′-ONH2 moiety to the 3′-ends of an oligonucleotide primer using 3′-deoxynucleoside triphosphates that have, instead of a 3′-OH moiety, a 3′-ONH2 moiety, where the nucleotides contain both standard and non-standard nucleobases, and where as a key claim limitation, substantially no hydroxylamine is present in the solutions used in the claimed processes.
Claims17
A process that adds to an oligonucleotide primer a single nucleotide that has a 3′-ONH2 moiety instead of a 3′-OH moiety, wherein said process comprises contacting an oligonucleotide primer in an aqueous buffered solution with an enzyme having terminal transferase activity and a compound having the structure:
- or one of its ionized forms, wherein Z is either H, OH, or OCH3, B is a heterocycle selected from the group consisting of
- wherein Su indicates the point of attachment of the heterocycle to the sugar, R is either H, CH3, or a functionalized side chain, and X is N, and wherein the concentration of hydroxylamine in said aqueous buffered solution that is less than 1 micromolar.
Dependencies are inferred from the claim text.