Efficient product cleavage in template-free enzymatic synthesis of polynucleotides
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.
1 . A kit for enzymatically synthesizing a polynucleotide comprising an initiator attached to a support by a 5′ end and having a 3′-penultimate deoxyinosine and a 3′-terminal nucleotide having a free 3′-hydroxyl.
2 . The kit of claim 1 , wherein said support is a solid support.
3 . The kit of claim 1 , wherein the solid support is selected from the group consisting of a magnetic bead, a planar solid, and a membrane.
4 . The kit of claim 1 , further comprising a de-blocking agent.
5 . The kit of claim 1 , wherein the initiator is in a container.
6 . The kit of claim 3 , wherein the planar solid is a glass slide.