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US 7,615,625 B2Granted
In vitro amplification of nucleic acid molecules via circular replicons
AssigneeReplicon, Inc.
InventorJeffrey I. Auerbach
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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.
Claims23
A kit for exponentially amplifying, in vitro, a target polynucleotide region of a nucleic acid molecule, wherein the kit comprises one or more containers comprising:
- (A) a single-stranded first polynucleotide, wherein the polynucleotide (i) contains a polynucleotide region that is complementary in sequence to the target polynucleotide region of a nucleic acid molecule, and (ii) is a circular polynucleotide or is circularizable when hybridized to a polynucleotide comprising the target polynucleotide region;
- (B) a second polynucleotide, wherein the second polynucleotide is a primer having a 3′ terminus hybridized to the single-stranded first polynucleotide and comprises the target polynucleotide region;
- and
- (C) a third polynucleotide, wherein the third polynucleotide is a primer and comprises a polynucleotide region that is complementary in sequence to a region of the second polynucleotide,
- wherein, in the presence of a polymerase, the 3′ termini of the second and third polynucleotides are extended in a template-dependent manner to exponentially amplify the target polynucleotide region.
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Dependencies are inferred from the claim text.