Enhanced ligation reactions
AssigneeLife Technologies Corporation
InventorsStephen Hendricks, David King, Lei Xi, Marian Peris
Life of this patent
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Abstract
In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of at least one agent that generates a ligatable terminal 5′ phosphate group by removing an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, an aprataxin enzyme can catalyze removal of an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of an aprataxin enzyme under conditions suitable for ligating nucleic acid ends.
Claims14
A method for ligating nucleic acids comprising:
- (a) forming a first single-stranded nick on a nucleic acid duplex by hybridizing a polynucleotide to a first and second oligonucleotide, wherein the first and second oligonucleotides abut each other, and wherein one end of the polynucleotide, the first oligonucleotide or the second oligonucleotide is attached to a surface;
- (b) closing the first single-stranded nick by conducting a nucleic acid ligation reaction in the presence of a ligase and an agent that catalyzes removal of an adenylate group from a terminal 5′ phosphate of a nucleic acid.
A method for sequencing comprising:
- a) hybridizing a template polynucleotide to a first and second oligonucleotide probe so that the first and second oligonucleotide probes abut each other to form a nick, wherein the second oligonucleotide probe is labeled with a distinctive detectable reporter moiety and wherein the polynucleotide is attached to a surface;
- b) contacting the nick with at least one aprataxin enzyme and at least one ligase enzyme to close the nick;
- and c) detecting the distinctive detectable reporter moiety thereby determining the sequence of the second oligonucleotide probe hybridized to the template polynucleotide.
Dependencies are inferred from the claim text.