Abstract: The present invention is directed to terminal deoxynucleotidyltransferase (TdT) variants that (i) comprise an amino acid sequence that is at least a specified percent identical to an indicated SEQ ID NOs and have at least one substitution at Q455 or at least Q455 plus at least one further substitution at G186, S248, T331, Q390, K394 or H466 (where positions are with respect to SEQ ID NO 1 and functionally equivalent positions in indicated SEQ ID NOs), (ii) are capable of template-free extension of a polynucleotide, and (iii) exhibit enhanced stability or enhanced efficiency in incorporating 3?-0-blocked nucleoside triphosphates into a polynucleotide. The invention is also directed to the use of these TdT variants for synthesizing polynucleotides of any predetermined sequence.

Abstract: The invention is directed to methods for synthesizing oligonucleotides directly on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3?-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3? hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.

Abstract: The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides that include or enable a step of efficiently cleaving the polynucleotide products from its initiator using an endonuclease V activity and initiator with a 3?-penultimate deoxyinosine.

Abstract: Methods for preparing 3?-O-amino-2?-deoxyribonucleoside-5?-triphosphates with reduced 3?-hydroxy-2?-deoxyribonucleoside-5?-triphosphate contamination by converting 3?-(N-acetone-oxime)-2?-deoxynucleoside triphosphate to 3?-O-amine-2?-deoxynucleoside triphosphate by treatment with an aryl-oxyamine and compositions produced therefrom.

Abstract: The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides that include or enable a step of efficiently cleaving the polynucleotide products from its initiator using an endonuclease V activity and initiator with a 3?-penultimate deoxyinosine.

Abstract: The invention relates to a process for preparing a 3?-O-amino-ribonucleotide. It also relates to a compound of formula (III), and its use as a precursor for the synthesis of a 3?-O-amino-ribonucleotide. In formula (III), B is a nitrogenous base or a protected derivative thereof, G and G? are identical or different, and are a protecting group, and RN— is a phthalimido or an imino group.

Abstract: The invention is directed to methods for synthesizing oligonucleotides direction on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3?-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3? hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.

Abstract: The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides that include or enable a step of efficiently cleaving the polynucleotide products from its initiator using endonuclease V activity and initiator with a 3?-penultimate deoxyinosine.

Abstract: The invention relates to a process for preparing a 3?-O-amino-ribonucleotide. It also relates to a compound of formula (III), and its use as a precursor for the synthesis of a 3?-O-amino-ribonucleotide. In formula (III), B is a nitrogenous base or a protected derivative thereof, G and G? are identical or different, and are a protecting group, and RN— is a phthalimido or an imino group.

Abstract: The invention is directed to methods for synthesizing oligonucleotides directly on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3?-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3? hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.

Abstract: The present invention relates to variants of Terminal deoxynucleotidyl Transferase (TdT), each of which (i) has an amino acid sequence similarity to SEQ ID NO: 2. 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33 or 35 with corresponding amino acid substitutions, (ii) is capable of synthesizing a nucleic acid fragment without a template and (iii) is capable of incorporating a modified nucleotide into the nucleic acid fragment.

Abstract: Methods for preparing 3?-O-amino-2?-deoxyribonucleoside-5?-triphosphates with reduced 3?-hydroxy-2?-deoxyribonucleoside-5?-triphosphate contamination by converting 3?-(N-acetone-oxime)-2?-deoxynucleoside triphosphate to 3?-O-amine-2?-deoxynucleoside triphosphate by treatment with an aryl-oxyamine and compositions produced therefrom.

Abstract: The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides that include or enable a step of efficiently cleaving the polynucleotide products from its initiator using endonuclease V activity and initiator with a 3?-penultimate deoxyinosine.

Abstract: The invention is directed to methods for synthesizing oligonucleotides direction on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3?-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3? hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.

Abstract: The invention is directed to methods for synthesizing oligonucleotides direction on biomolecules or cells living or fixed. In some embodiments, template-free enzymatic synthesis is implemented under biological conditions with successive cycles of (i) enzymatic addition of a 3?-O-blocked nucleoside triphosphate and (ii) enzymatic deblocking of the incorporated nucleotide to regenerate a free 3? hydroxyl. The invention has applications in single-cell cDNA library construction and analysis.