Abstract: The invention relates to compositions for generating a signal indicative of the presence of a target nucleic acid in a sample, where the compositions include a probe having a 5? region and a 3? flap and a P. furiosus polymerase having 3? exonuclease activity.

Abstract: The present invention is directed to methods of generating a signal indicative of the presence of a target nucleic acid in a sample by forming a cleavage structure comprising duplex and single-stranded nucleic acid by incubating a sample comprising a target nucleic acid with a thermostable nucleic acid polymerase substantially lacking 5? to 3? exonuclease activity and cleaving said cleavage structure with a thermostable Fen nuclease lacking 5? to 3? synthetic activity to generate a signal.

Abstract: The present invention provides methods, kits and compositions for the detection of an analyte. In the methods of the invention, a binding molecule coupled to a first polymerase is incubated with a modified polynucleotide template to form a copy of the modified polynucleotide template without any modified nucleotides. The unmodified copy is detected in a second amplification/primer extension reaction using a second polymerase that is unable to amplify the modified polynucleotide template. Detection of the unmodified copy is indicative of the presence and/or amount of the analyte in the sample. In place of the binding molecule coupled to a first polymerase, a pair of analyte-specific probes can also be used. The first analyte specific probe comprises a first binding moiety and a first portion of a first polymerase and the second analyte specific probe comprises a second binding moiety and a second portion of the first polymerase.

Abstract: The invention provides methods, compositions and kits for generating a signal indicative of the presence of a target nucleic acid sequence in a sample comprising forming a cleavage structure by incubating a sample comprising a target nucleic acid sequence with upstream and downstream oligonucleotides, and cleaving the cleavage structure with a nuclease to generate a signal. The presence of a detectable signal is indicative of the presence of a target nucleic acid sequence and a non-invasive cleavage structure.

Abstract: The present invention provides methods, kits and compositions for the detection of an analyte. In the methods of the invention, a complex is formed between two or more analyte specific probes (ASP) and an analyte. The analyte specific probes each have a portion of a polymerase which interact to form a functional polymerase complex upon binding of the ASP to the analyte. The functional polymerase complex then generates a detectable signal which is indicative of the presence and/or amount of the analyte in the sample.

Abstract: The present invention is directed to kits and compositions for generating a signal indicative of the presence of a target nucleic acid sequence in a sample, comprising a nucleic acid polymerase lacking 5? to 3? exonuclease activity and a thermostable FEN nuclease consisting of a 5? to 3? exonuclease and/or endonuclease activity.

Abstract: The invention provides compositions, kits and methods of generating a signal indicative of the presence of a target nucleic acid sequence in a sample by forming a cleavage structure. The cleavage structure is formed by incubating a sample containing a target nucleic acid with a downstream probe that forms a 3? flap when hybridized to the target. The cleavage structure is cleaved with a 3? nuclease and a detectable signal is produced.

Abstract: The invention relates to compositions and methods for generating a signal indicative of the presence of a target nucleic acid in a sample utilizing a primer-probe duplex.

Abstract: The invention relates to compositions and methods for generating a signal indicative of the presence of a target nucleic acid in a sample utilizing a cleavage resistant probe.

Abstract: The invention relates to compositions and methods for generating a signal indicative of the presence of a target nucleic acid in a sample, where the compositions and methods include an RNA polymerase, a FEN nuclease, and a probe.

Abstract: The invention provides cells and methods of circularizing linear DNA molecules. The cell is an isolated Escherichia coli cell which transiently expresses the Cre recombinase protein from an integrated Cre recombinase gene, and which is at least transiently repressed for RecBCD activity. The cells are used in a method of circularizing a linear DNA molecule comprising at least two loxP sites. The DNA molecule is introduced into the cells, and the linear DNA molecule is joined at said loxp sites.

Abstract: The invention relates to compositions and methods for detection of nucleic acid sequences. The invention further relates to kit format of said compositions for detection of nucleic acid sequences. Oligonucleotide probes of the invention comprise a target binding sequence and a sequence at least partially complementary thereto, joined by an optional linker to form a hairpin structure in the absence of the target nucleic acid sequence. The probes of the invention comprise a pair of moieties that produce a detectable signal when the probe hybridizes to the target sequence.

Abstract: The present invention provides methods, kits and compositions for the detection of an analyte. In the methods of the invention, a complex is formed between two or more analyte specific probes (ASP) and an analyte. The analyte specific probes each have a portion of a polymerase which interact to form a functional polymerase complex upon binding of the ASP to the analyte. The functional polymerase complex then generates a detectable signal which is indicative of the presence and/or amount of the analyte in the sample.

Abstract: The invention relates to a method of generating a signal indicative of the presence of a target nucleic acid in a sample, where the method includes forming a cleavage structure by incubating a sample comprising a target nucleic acid with a probe having a secondary structure that changes upon binding of the probe to the target nucleic acid and further comprising a binding moiety. The invention also includes the steps of cleaving the cleavage structure with a nuclease to release a nucleic acid fragment to generate a signal, wherein generation of the signal is indicative of the presence of a target nucleic acid in a sample, and detecting and/or measuring the amount of the fragment captured by binding of a binding moiety to a capture element on a solid support.

Abstract: The invention relates to and methods for generating a signal indicative of the presence of a target nucleic acid in a sample. The compositions and methods include a reverse transcriptase, a nuclease, an upstream primer and downstream probe.

Abstract: The invention relates to compositions for generating a signal indicative of the presence of a target nucleic acid in a sample, where the compositions include an RNA polymerase, a FEN nuclease, and a probe.

Abstract: The invention provides methods, compositions and kits for generating a signal indicative of the presence of a target nucleic acid sequence in a sample comprising forming a cleavage structure by incubating a sample comprising a target nucleic acid sequence with upstream and downstream oligonucleotides, and cleaving the cleavage structure with a nuclease to generate a signal. The presence of a detectable signal is indicative of the presence of a target nucleic acid sequence and a non-invasive cleavage structure.

Abstract: The invention features a novel isolated Family B DNA polymerase, a Thermococcus polymerase JDF-3, and mutant recombinant forms thereof. Mutant polymerases of the invention are deficient in 3? to 5? exonuclease activity and/or exhibit reduced discrimination against non-conventional nucleotides relative to the wild-type form of the polymerase.

Abstract: The invention features a novel isolated Family B DNA polymerase, a Thermococcus polymerase JDF-3, and mutant recombinant forms thereof. Mutant polymerases of the invention are deficient in 3? to 5? exonuclease activity and/or exhibit reduced discrimination against non-conventional nucleotides relative to the wild-type form of the polymerase.

Abstract: The present invention provides methods, nucleic acids, compositions, and kits for detecting microRNA (miRNA) in samples. The methods comprise designing mRNA-specific primers, adding a polyA tail to the miRNA, and using reverse transcription and amplification to detect the miRNA. The nucleic acids, compositions, and kits typically comprise some or all of the components necessary to practice the methods of the invention.