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Abstract

Disclosed are compositions and methods for proximity-mediated rolling circle amplification and for real-time detection of proximity-mediated rolling circle amplification products. Rolling circle amplification (RCA) refers to nucleic acid amplification reactions involving replication of a circular nucleic acid template to form a long strand with tandem repeats of the sequence complementary to the circular template. In proximity-mediated RCA, binding guide conjugates are brought into close proximity, generally by associating them to the same analyte or to two analytes in close proximity. The binding guide conjugates comprise a specific binding molecule and a guide oligonucleotide. The guide oligonucleotides are complementary to guide complement portions on half circle probes. The complementary sequences between the guide oligonucleotides and half circle probes allow both ends of each half circle probe to hybridize adjacent to an end of the other half circle probe and to be ligated together to form a circular nucleic acid molecule comprising the two half circle probes. This circular nucleic acid molecule can then be used as the template in RCA.

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First Claim