An authentication assay using embedded deoxyribonucleic acid (DNA) taggants includes a substrate and a sample of an authenticity label collected from a product. The substrate has a plurality of assay locations, each of which includes a reporter oligonucleotide bound to the substrate. The reporter oligonucleotide includes a first region with a single-stranded toehold sequence, a second region with a universal sequence, and a third region with a unique sequence, the second and third regions being prehybridized with a complementary strand. The sample includes at least one fluorophore-labeled DNA taggant complementary to the first and second regions of the reporter oligonucleotide. Incubation of the substrate with the sample results in a toehold-mediated DNA strand displacement reaction that exchanges the complementary strand for the fluorophore-labeled DNA taggant. Excitation of the fluorophore molecule attached to the DNA taggant produces a pattern of light emitted at one or more assay locations.

An authentication assay using embedded deoxyribonucleic acid (DNA) taggants includes a substrate and a sample of an authenticity label collected from a product. The substrate has a plurality of assay locations, each of which includes a reporter oligonucleotide bound to the substrate. The reporter oligonucleotide includes a first region with a single-stranded toehold sequence, a second region with a universal sequence, and a third region with a unique sequence, the second and third regions being prehybridized with a complementary strand. The sample includes at least one fluorophore-labeled DNA taggant complementary to the first and second regions of the reporter oligonucleotide. Incubation of the substrate with the sample results in a toehold-mediated DNA strand displacement reaction that exchanges the complementary strand for the fluorophore-labeled DNA taggant. Excitation of the fluorophore molecule attached to the DNA taggant produces a pattern of light emitted at one or more assay locations.

Provided herein are compositions, kits and methods for the production of strand-specific cDNA libraries. The compositions, kits and methods utilize properties of double stranded polynucleotides, such as RNA-cDNA duplexes to capture and incorporate a novel sequencing adapter. The methods are useful transcriptome profiling by massive parallel sequence, such as full-length RNA sequencing (RNA-Seq) and 3′ tag digital gene expression (DGE).

Provided herein are compositions, kits and methods for the production of strand-specific cDNA libraries. The compositions, kits and methods utilize properties of double stranded polynucleotides, such as RNA-cDNA duplexes to capture and incorporate a novel sequencing adapter. The methods are useful transcriptome profiling by massive parallel sequence, such as full-length RNA sequencing (RNA-Seq) and 3′ tag digital gene expression (DGE).

The present invention provides a complex of LNA probe and graphene oxide, and a nucleic acid detection method using the same. In the present invention, LNA-containing molecular beacon is conjugated through covalent bonding with graphene oxide, a single strand of the molecular beacon binds to a target nucleic acid to form a complex, and the complex is separated from graphene oxide to induce a fluorescence signal. The molecular beacon and graphene oxide can be covalently bonded to minimize non-specific signals, and a LNA-added molecular beacon is designed in a double strand to detect a very low concentration of target nucleic acid with high sensitivity, as well as a fluorescent signal, and the multiple target nucleic acids can be detected simultaneously through diversification of the fluorescent signal to enable easy and accurate detection of a nucleic acid biomarker whose specific expression level is specifically changed according to diseases and disease progression.

Disclosed is a method of performing a non-isothermal nucleic acid amplification reaction, the method comprising the steps of: (a) mixing a target sequence with one or more complementary single stranded primers in conditions which permit a hybridisation event in which the primers hybridise to the target, which hybridisation event, directly or indirectly, leads to the formation of a duplex structure comprising two nicking sites disposed at or near opposite ends of the duplex; and performing an amplification process by; (b) causing a nick at each of said nicking sites in the strands of the duplex; (c) using a polymerase to extend the nicked strands so as to form newly synthesised nucleic acid, which extension with the polymerase recreates nicking sites; (d) repeating steps (b) and (c) as desired so as to cause the production of multiple copies of the newly synthesised nucleic acid; characterised in that the temperature at which the method is performed is non-isothermal, and subject to a reduction of at least 2° C. during the amplification process of steps (b)-(d).

Provided herein are compositions, kits and methods for the production of strand-specific cDNA libraries. The compositions, kits and methods utilize properties of double stranded polynucleotides, such as RNA-cDNA duplexes to capture and incorporate a novel sequencing adapter. The methods are useful transcriptome profiling by massive parallel sequence, such as full-length RNA sequencing (RNA-Seq) and 3′ tag digital gene expression (DGE).

Provided herein are compositions, kits and methods for the production of strand-specific cDNA libraries. The compositions, kits and methods utilize properties of double stranded polynucleotides, such as RNA-cDNA duplexes to capture and incorporate a novel sequencing adapter. The methods are useful transcriptome profiling by massive parallel sequence, such as full-length RNA sequencing (RNA-Seq) and 3′ tag digital gene expression (DGE).

Every year, approximately 94 million cases of Salmonella gastroenteritis, with 155000 deaths, are reported each year and 85% of them reported to be food-borne. Investigation of the foods whether they are clean for Salmonella and sensitivity, easy applicability, absence of false positivity and negativity and the speed are the features sought in the analysis method for this investigation. It is not desirable for analysis to detect the presence of dead bacteria in food. Although the final product does not contain microbiologically harmful live bacteria during the food process, the detection of dead bacteria transmitted before the process causes the food product to be unfairly diagnosed as harmful. To prevent this situation, the analysis kits depending on molecular methods, increase their microorganism detection levels up to to 10.sup.4 while reducing their sensitivity. Since the molecular methods cannot discriminate dead and live organisms, a confirmation test is required to prove that the positive result of the analysis belongs to the live bacteria in the food, which results in additional cost and time loss. In the same way, it is necessary to verify whether the colonies that grow in the gold standard culture method, belong to Salmonella bacteria. In the developed system; 10.sup.5 dead bacterial DNA is eliminated in the food to prevent false positive results and the minimum detection limit is 10 bacteria. Also, in developed system, 4 primers specific to 6 regions of DNA are used. Therefore, the specificity of the method is very high (99.9%) and no verification test is needed. Since PCR systems require a device with complex temperature control units, they can make analysis in a laboratory-dependent manner. In the proposed system, DNA is amplified at constant temperature; no temperature cycle is required, therefore no complex instrument and laboratory infrastructure are required. All the procedures can be easily performed outside the laboratory on a portable mini-heater where pre-enrichment, DNA isolation from the sample and PCR steps are performed. For molecular analyses, the device is required to display the result of imaging or analysis. In the developed method, DNAs amplified by the loop-mediated isothermal DNA amplification method, are hybridized and combined with the labeled probe and then can be read by lateral flow method with the naked eye. As the results are visible by eye, no additional device is required. The classical culture method is accepted as the gold standard, but the duration of analysis is 7 days for positive samples, 3 days with verification test, for the molecular methods, and 5.5 hours including pre-enrichment time

Every year, approximately 94 million cases of Salmonella gastroenteritis, with 155000 deaths, are reported each year and 85% of them reported to be food-borne. Investigation of the foods whether they are clean for Salmonella and sensitivity, easy applicability, absence of false positivity and negativity and the speed are the features sought in the analysis method for this investigation. It is not desirable for analysis to detect the presence of dead bacteria in food. Although the final product does not contain microbiologically harmful live bacteria during the food process, the detection of dead bacteria transmitted before the process causes the food product to be unfairly diagnosed as harmful. To prevent this situation, the analysis kits depending on molecular methods, increase their microorganism detection levels up to to 10.sup.4 while reducing their sensitivity. Since the molecular methods cannot discriminate dead and live organisms, a confirmation test is required to prove that the positive result of the analysis belongs to the live bacteria in the food, which results in additional cost and time loss. In the same way, it is necessary to verify whether the colonies that grow in the gold standard culture method, belong to Salmonella bacteria. In the developed system; 10.sup.5 dead bacterial DNA is eliminated in the food to prevent false positive results and the minimum detection limit is 10 bacteria. Also, in developed system, 4 primers specific to 6 regions of DNA are used. Therefore, the specificity of the method is very high (99.9%) and no verification test is needed. Since PCR systems require a device with complex temperature control units, they can make analysis in a laboratory-dependent manner. In the proposed system, DNA is amplified at constant temperature; no temperature cycle is required, therefore no complex instrument and laboratory infrastructure are required. All the procedures can be easily performed outside the laboratory on a portable mini-heater where pre-enrichment, DNA isolation from the sample and PCR steps are performed. For molecular analyses, the device is required to display the result of imaging or analysis. In the developed method, DNAs amplified by the loop-mediated isothermal DNA amplification method, are hybridized and combined with the labeled probe and then can be read by lateral flow method with the naked eye. As the results are visible by eye, no additional device is required. The classical culture method is accepted as the gold standard, but the duration of analysis is 7 days for positive samples, 3 days with verification test, for the molecular methods, and 5.5 hours including pre-enrichment time