Use of deoxyribose nicotinamide adenine dinucleotide to enhance the specificity of NAD.sup.+ -dependent ligation reactions
AssigneeBio-Rad Laboratories, Inc.
InventorsRobert B. Wallace, Hector Juarez-Salinas, Luis Ugozzoli
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Abstract
This invention provides methods and compounds relating to the use of deoxyribose nicotinamide adenine dinucleotide (dNAD.sup.+) analogues in NAD.sup.+ -dependent ligation reactions. The compounds and methods may be used in NAD.sup.+ -dependent ligation reactions generally, and to increase the specificity of NAD.sup.+ -dependent ligation reactions.
Claims20
A method for ligating nucleic acid oligonucleotides, comprising the steps of:
- (a) providing a ligation reaction mixture comprising a nicotinamide adenine dinucleotide-dependent nucleic acid ligase, nucleic acid oligonucleotides, and a deoxyribose nicotinamide adenine dinucleotide, and
- (b) incubating said mixture under conditions such that said oligonucleotides are ligated to one another.
A method for specifically ligating oligonucleotides complementary to a target polynucleotide which comprises the steps of combining in an aqueous ligation solution the target polynucleotide, nucleic acid oligonucleotides complementary to adjacent portions a deoxyribose nicotinamide adenine dinucleotide, not the target, a nicotinamide adenine dinucleotide-dependant nucleic acid ligase capable of ligating the oligonucleotides, Mg.sup.++ and ATP;
- and incubating the ligation mixture to permit the nucleic acid ligase to ligate nucleic acid oligonucleotides associated with the target nucleic acid.
A kit for ligating DNA oligonucleotides comprising a container containing an nicotinamide adenine dinucleotide-dependant DNA ligase and deoxyribose nicotinamide adenine dinucleotide.
Dependencies are inferred from the claim text.