Amplification of nucleic acid molecules via circular replicons
AssigneeReplicon, Inc.
InventorJeffrey I. Auerbach
Life of this patent
Estimating the term…
Abstract
Methods and compositions suitable for accomplishing the in vitro amplification of nucleic acid molecules via enzymatic means are provided. The preferred means employ circular rather than linear replicons. Means for producing such circular replicons from linear reactants are also provided.
Claims19
A method for amplifying a target polynucleotide region of an initial linear nucleic acid molecule, which method comprises the steps of:
- (A) providing a partially single-stranded, partially double-stranded nucleic acid complex, wherein a first strand of said molecule contains a polynucleotide region that is complementary in sequence to said target polynucleotide region and is a circular strand lacking termini and a second strand of said complex is non-circular and has a 3′ terminus hybridized to said circular strand of said molecule;
- and
- (B) incubating said partially single-stranded, partially double-stranded nucleic acid molecule in the presence of a polymerase and under conditions sufficient to permit said polymerase to extend said 3′ terminus of said non-circular strand in a template-dependent manner using said circular strand of said molecule as said template to thereby produce an extension product;
- wherein said template-dependent extension of said 3′ terminus of said non-circular strand causes strand displacement of a portion of said non-circular strand and results in said amplification of said target polynucleotide region.
An in vitro polynucleotide complex comprising a first, second and third strand of DNA, wherein said first strand is circular and lacks termini and said second and third strands are non-circular and possess termini;
- wherein a 3′ terminal region of said second strand is complementary to a region of said circular first strand, and is hybridized thereto, and a non-3′ terminal region of said second strand is complementary to a region of said third strand, and is hybridized thereto.
An in vitro polynucleotide complex comprising at least a first, a second, and a third strand of DNA, wherein:
- said first strand is circular and lacks termini;
- said second strand is non-circular, and possesses a 3′ terminal region that is complementary to a region of said circular first strand, and is hybridized thereto;
- and
- said third strand is non-circular, and possesses a 3′ terminal region that is complementary to a region of said second strand and is hybridized thereto.
Dependencies are inferred from the claim text.