Patent ID: 11905541
Assignee: DNA SCRIPT SAS
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method of synthesizing a polynucleotide having a predetermined sequence, the method comprising the steps of:
a) providing an initiator having a 3′-penultimate deoxyinosine and a 3′-terminal nucleotide having a free 3′-hydroxyl;
b) repeating cycles of (i) contacting under elongation conditions the initiator or elongated fragments having free 3′-O-hydroxyls with a 3′-O-blocked nucleoside triphosphate and a template-independent DNA polymerase so that the initiator or elongated fragments are elongated by incorporation of the 3′-O-blocked nucleoside triphosphate to form 3′-O-blocked elongated fragments, and (ii) deblocking the elongated fragments to form elongated fragments having free 3′-hydroxyls, until the polynucleotide is formed, wherein said template-independent DNA polymerase is a terminal deoxynucleotidyl transferase; and
c) treating the polynucleotide with an endonuclease V activity to cleave the polynucleotide from the initiator, wherein said endonuclease V activity is provided by a prokaryotic endonuclease V.