Abstract: A hot start enzyme composition is described that includes a hot start nuclease, a nucleic acid polymerase, and a substantially double-stranded oligonucleotide that inhibits the catalytic activity of the nucleic acid polymerase at temperatures lower than the melting temperature of the oligonucleotide.

Abstract: A reversibly modified ‘hot start’ RNase H enzyme composition is described for the improved CATACLEAVE™ probe detection of nucleic acid sequences in a test sample. A key feature of the enzyme composition is the ability to regulate the catalytic activity of the RNase H during the course of a reverse transcription-PCR cycle. Thus, RNase H activity can be initially suppressed to minimize degradation of RNA:DNA primer heteroduplexes prior to reverse transcription. After cDNA synthesis is complete, RNase H activity is induced to promote the cleavage and fluorescent detection of CATACLEAVE™ probes that anneal to target DNA sequences within the reverse transcriptase-PCR products. The inducible RNase H enzyme is amenable to high throughput applications requiring one step reverse transcriptase CATACLEAVE™ PCR in a single reaction mix.

Abstract: Disclosed is a method in the fields of biochemistry and molecular biology. The method is related to improve cleavage kinetics of labeled oligonucleotide probes and, consequently, increases signal-to-noise ratio in detecting nucleic acids.

Abstract: A method is described for the real-time detection of HIV-1 gene target DNA in a sample, including obtaining HIV-1 specific cDNA by reverse transcription, amplifying a portion of the cDNA, and detecting the amplicons so produced using probe labeled with a detectable marker.

Abstract: A reversibly modified ‘hot start’ RNase H enzyme composition is described for the improved CATACLEAVE™ probe detection of nucleic acid sequences in a test sample. A key feature of the enzyme composition is the ability to regulate the catalytic activity of the RNase H during the course of a reverse transcription-PCR cycle. Thus, RNase H activity can be initially suppressed to minimize degradation of RNA:DNA primer heteroduplexes prior to reverse transcription. After cDNA synthesis is complete, RNase H activity is induced to promote the cleavage and fluorescent detection of CATACLEAVE™ probes that anneal to target DNA sequences within the reverse transcriptase-PCR products. The inducible RNase H enzyme is amenable to high throughput applications requiring one step reverse transcriptase CATACLEAVE™ PCR in a single reaction mix.

Abstract: A reversibly modified ‘hot start’ RNAse H enzyme composition is described for the improved CATACLEAVE™ probe detection of nucleic acid sequences in a test sample. A key feature of the enzyme composition is the ability to regulate the catalytic activity of the RNAse H during the course of a reverse transcription-PCR cycle. Thus, RNAse H activity can be initially suppressed to minimize degradation of RNA:DNA primer heteroduplexes prior to reverse transcription. After cDNA synthesis is complete, RNAse H activity is induced to promote the cleavage and fluorescent detection of CATACLEAVE™ probes that anneal to target DNA sequences within the reverse transcriptase-PCR products. The inducible RNAse H enzyme is amenable to high throughput applications requiring one step reverse transcriptase CATACLEAVE™ PCR in a single reaction mix.

Abstract: A hot start enzyme composition is described that includes a hot start nuclease, a nucleic acid polymerase, and a substantially double-stranded oligonucleotide that inhibits the catalytic activity of the nucleic acid polymerase at temperatures lower than the melting temperature of the oligonucleotide.

Abstract: Disclosed is a method in the fields of biochemistry and molecular biology. The method is related to improve cleavage kinetics of labeled oligonucleotide probes and, consequently, increases signal-to-noise ratio in detecting nucleic acids.

Abstract: A method is described for the real-time detection of Salmonella species in foods and on surfaces. Salmonella are enriched in media to increase their cell density prior to analysis. DNA is recovered by lysis in the presence of azide, proteinase K, and detergent. Real-time detection of Salmonella species is performed in a PCR reaction using gene specific primers and a cleavable chimeric fluorescent probe. The method also describes an internal control to confirm the efficiency of nucleic acid amplification and detection. The method is amenable to medium and high throughput analysis.

Abstract: Disclosed are methods and kits for the detection of a polymorphism during real-time PCR. Real-time PCR amplification of a target nucleic acid sequence is performed using PCR primer primers that anneal to sequences flanking a single nucleotide polymorphism (SNP) of interest. The real-time PCR reaction includes a labeled probe comprising a RNA sequence that is designed to anneal to DNA sequences at the location of the SNP. An RNA:DNA heteroduplex can then form between the SNP in the PCR fragment and the probe's RNA sequences that are complementary to the SNP. RNase H cleavage of the RNA sequence in the RNA:DNA heteroduplex results in increase in intensity of the signal generated from the label that is indicative of the presence of an SNP in the target nucleic acid.

Abstract: A method is described for the real-time detection of HIV-1 gene target DNA in a sample, including obtaining HIV-1 specific cDNA by reverse transcription, amplifying a portion of the cDNA, and detecting the amplicons so produced using probe labeled with a detectable marker.

Abstract: Oligonucleotides, a kit, and a method for detecting E. coli O157:H7 strains are provided. According to the kit for detecting E. coli O157:H7 strains and the method of detecting E. coli O157:H7 strains by using the kit, the results of the detection can be rapidly identified with a reduced number of copies of a sample in real-time.

Abstract: A kit for detecting HCV in a test sample is disclosed. In addition a method is described for the real-time detection of HCV in a test sample using the kit. According to method of detection, the results of the detection can be rapidly identified with a reduced number of copies of a sample in real-time.

Abstract: A reversibly modified ‘hot start’ RNAse H enzyme composition is described for the improved CATACLEAVE™ probe detection of nucleic acid sequences in a test sample. A key feature of the enzyme composition is the ability to regulate the catalytic activity of the RNAse H during the course of a reverse transcription-PCR cycle. Thus, RNAse H activity can be initially suppressed to minimize degradation of RNA:DNA primer heteroduplexes prior to reverse transcription. After cDNA synthesis is complete, RNAse H activity is induced to promote the cleavage and fluorescent detection of CATACLEAVE™ probes that anneal to target DNA sequences within the reverse transcriptase-PCR products. The inducible RNAse H enzyme is amenable to high throughput applications requiring one step reverse transcriptase CATACLEAVE™ PCR in a single reaction mix.

Abstract: The invention teaches a novel reagent formulation for the efficient preparation of a nucleic acid template from cells for high throughput real-time PCR analysis. The reagent permits rapid cell lysis and template preparation without the need for template purification and isolation. The reagent therefore dramatically improves throughput of real-time PCR analysis while at the same time permitting the rapid and sensitive real-time Catacleave PCR detection of a single molecule of nucleic acid template in as little as about 35 cycles of PCR amplification.

Abstract: A method is described for the real-time detection of Salmonella species in foods and on surfaces. Salmonella are enriched in media to increase their cell density prior to analysis. DNA is recovered by lysis in the presence of azide, proteinase K, and detergent. Real-time detection of Salmonella species is performed in a PCR reaction using gene specific primers and a cleavable chimeric fluorescent probe. The method also describes an internal control to confirm the efficiency of nucleic acid amplification and detection. The method is amenable to medium and high throughput analysis.