Low Sequence Bias Single-Stranded DNA Ligation
AssigneeThe Penn State Research Foundation
InventorsPhilip C. Bevilacqua, Sarah M. Assmann, Yiliang Ding, Chun Kit Kwok
Life of this patent
Estimating the term…
Abstract
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.
Claims20
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.
A composition comprising a donor nucleic acid molecule, wherein the molecule comprises a stem-loop structure and a 3′ overhang, further wherein the molecule comprises a continuous primer binding site that encompasses a portion of the stem and a portion of the loop structure.
A kit comprising a donor nucleic acid molecule, wherein the molecule comprises a stem-loop structure and a 3′ overhang, further wherein the molecule comprises a continuous primer binding site that encompasses a portion of the stem and a portion of the loop structure.
Dependencies are inferred from the claim text.