Novel use
AssigneeNuclera Nucleics Ltd.
InventorsMichael C. CHEN, Radu A. LAZAR, Jiahao HUANG, Gordon R. MCINROY
Life of this patent
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Abstract
The invention relates to the use of specific terminal deoxynucleotidyl transferase (TdT) enzymes in a method of nucleic acid synthesis, to methods of synthesizing nucleic acids, and to the use of kits comprising said enzymes in a method of nucleic acid synthesis. The invention also relates to the use of terminal deoxynucleotidyl transferases and 3′-blocked nucleotide triphosphates in a method of template independent nucleic acid synthesis.
Claims29
A method of nucleic acid synthesis comprising a step of providing a terminal deoxynucleotidyl transferase (TdT) enzyme comprising an amino acid sequence having at least 95% sequence identity to any one of SEQ ID NOs:
- 1 to 5 and 8, such as any of SEQ ID NOs:
- 1, 2, or 8.
(canceled)
9-10. (canceled)
12-13. (canceled)
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17-19. (canceled)
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24-25. (canceled)
A method of template-independent acid synthesis comprising a step of providing a 3′-blocked nucleotide triphosphate, wherein the 3′-blocked nucleotide triphosphate is selected from a compound of formula (I), (II), (III), or (IV):
- wherein
- R1 represents NRaRb, wherein Ra and Rb independently represent hydrogen or C1-6 alkyl,
- R2 represents hydrogen, C1-6 alkyl, C1-6 alkoxy, COH or COOH,
- X represents C1-6 alkyl, NH2, N3 or —OR3,
- R3 represents C1-6 alkyl, CH2N3, NH2 or allyl,
- Y represents hydrogen, halogen or hydroxyl, and
- Z represents CR4 or N, wherein R4 represents hydrogen, C1-6 alkyl, C1-6 alkoxy, COH or COOH.
30-34. (canceled)
A method of nucleic acid synthesis comprising a step of providing an inorganic pyrophosphatase such as purified, recombinant inorganic pyrophosphatase from Saccharomyces cerevisiae.
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Dependencies are inferred from the claim text.