Patent ID: 8809022

Claim:
A method for enzymatically ligating a pre-adenylated donor molecule to RNA, the method comprising reacting the pre-adenylated donor molecule and the 3′ hydroxyl group of a 5′ phosphorylated or de-phosphorylated RNA in the absence of adenosine triphosphate and in the presence of an enzyme comprising a truncated T4 RNA ligase 2 (1-249), wherein said enzyme includes a substitution of an arginine with a lysine at a position corresponding to position 55 as encoded by SEQ ID. NO: 3, and wherein the truncated T4 RNA ligase 2 is capable of ligating the pre-adenylated donor molecule to the 3′ hydroxyl group of the optionally de-phosphorylated RNA in the absence of adenosine triphosphate.