Ligation method employing eukaryotic tRNA ligase

US8927245B2 · kind B2 · utility

Assignee

• AGILENT TECHNOLOGIES, INC. · US

Inventors

• Gusti Zeiner · Pacifica, US
• Robert A. Ach · San Francisco, US

Key dates

• Filing date: Dec 15, 2011
• Grant date: Jan 6, 2015
• Expiry date: Feb 1, 2033

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/1096
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Provided herein is a method of preparing an RNA sample comprising: a) obtaining an RNA sample comprising: i. long RNA molecules that may be unfragmented or fragmented to contain 5′-OH group and a 2′-3′-cyclic phosphate group; and ii. short RNA molecules that comprise a 5′ phosphate group and a 3′ OH group; and b) contacting the RNA sample with an adaptor comprising either a 2′-PO group and 3′-OH group or a 2′,3′-cyclic phosphate group in the presence of a eukaryotic tRNA ligase, thereby producing a ligated RNA sample in which a) the short RNA molecules are selectively ligated to the adaptor or b) the short RNA molecules and long RNA fragments are selectively ligated to the adaptor.