Low Sequence Bias Single-Stranded DNA Ligation

摘要

The invention provides compositions and methods for ligating single stranded nucleic acids wherein the ligation is based on fast, efficient, and low-sequence bias hybridization of an acceptor molecule with a donor molecule. In one embodiment, the structure of the donor molecule comprises a stem-loop intramolecular nucleotide base pairing (i.e., hairpin) and a 3′-overhang region such that the overhang is able to hybridize to nucleotides present in the 3′ end of the acceptor molecule.

主权项

1. A method of producing a ligated single stranded nucleic acid molecule, the method comprising: a) contacting a single-stranded acceptor nucleic acid molecule with a donor nucleic acid molecule, wherein the donor nucleic acid molecule comprises one or more nucleic acids having a stem region and a single-stranded 3′ terminal overhang region; b) hybridizing the single stranded 3′ terminal overhang region of the donor nucleic acid molecule to the acceptor molecule thereby forming an acceptor-donor hybrid molecule comprising a nick or gap between the acceptor nucleic acid and donor nucleic acid molecule; c) and ligating the 5′ end of the donor nucleic acid molecule to the 3′ end of the acceptor nucleic acid molecule thereby generating a ligated product.