Disclosed is a method of performing a non-isothermal nucleic acid amplification reaction, wherein the temperature at which the method is performed is non-isothermal and subject to a reduction of at least 2° C. during amplification process steps. The present invention provides an improved nucleic acid amplification technique having one or more advantages over existing techniques including, for example, decreased reaction time, increased yield, and decreased non-specific amplification products.

Disclosed is a method of performing a non-isothermal nucleic acid amplification reaction, wherein the temperature at which the method is performed is non-isothermal and subject to a reduction of at least 2° C. during amplification process steps. The present invention provides an improved nucleic acid amplification technique having one or more advantages over existing techniques including, for example, decreased reaction time, increased yield, and decreased non-specific amplification products.

Disclosed are methods for detecting a target nucleic acid in a sample. The methods include contacting the sample, in the presence of a polymerase and an endonuclease, with a first oligonucleotide comprising, in the 5′ to 3′ direction, a first signal DNA generation sequence, an endonuclease recognition site, and a sequence complementary to the 3′ end of a target nucleic acid; a second oligonucleotide comprising, in the 5′ to 3′ direction, a second signal DNA generation sequence, an endonuclease recognition site, and a sequence that is homologous to the first signal DNA generation sequence of the first oligonucleotide; a third oligonucleotide comprising, in the 5′ to 3′ direction, a third signal DNA generation sequence, an endonuclease recognition site, and a sequence that is homologous to the second signal DNA generation sequence of the second oligonucleotide.

Disclosed are methods for detecting a target nucleic acid in a sample. The methods include contacting the sample, in the presence of a polymerase and an endonuclease, with a first oligonucleotide comprising, in the 5′ to 3′ direction, a first signal DNA generation sequence, an endonuclease recognition site, and a sequence complementary to the 3′ end of a target nucleic acid; a second oligonucleotide comprising, in the 5′ to 3′ direction, a second signal DNA generation sequence, an endonuclease recognition site, and a sequence that is homologous to the first signal DNA generation sequence of the first oligonucleotide; a third oligonucleotide comprising, in the 5′ to 3′ direction, a third signal DNA generation sequence, an endonuclease recognition site, and a sequence that is homologous to the second signal DNA generation sequence of the second oligonucleotide.

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 hybridization event in which the primers hybridize to the target, which hybridization 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) using a nicking enzyme to cause 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 synthesized 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 synthesized nucleic acid.

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 hybridization event in which the primers hybridize to the target, which hybridization 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) using a nicking enzyme to cause 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 synthesized 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 synthesized nucleic acid.

The present application relates to a biosensor for detecting an analyte comprising a first and second working electrode; a detection probe functionalized on the first working electrode, the detection probe comprising a reporter moiety and a recognition moiety for an analyte; a capture probe functionalized on the second working electrode; and a counter electrode. Each working electrode is configured to provide a change in signal if the analyte is present. The biosensor can be used to detect an analyte in a sample.

The present application relates to a biosensor for detecting an analyte comprising a first and second working electrode; a detection probe functionalized on the first working electrode, the detection probe comprising a reporter moiety and a recognition moiety for an analyte; a capture probe functionalized on the second working electrode; and a counter electrode. Each working electrode is configured to provide a change in signal if the analyte is present. The biosensor can be used to detect an analyte in a sample.

Provided herein are methods and kits for parallel detection one or more target sequences across multiple samples, comprising separating a set of samples into one or more pooled sets, wherein each sample may comprise an initial amplicon comprising one or more target sequences and at least one barcode; conducting an amplification reaction on the one or more pooled sets to further amplify the amplicons, and optionally further adding an additional barcode to the amplicon; sequencing the amplicons; identifying individual samples from the pooled sample set that are positive for the one or more target sequences based on sequencing of the amplicons, wherein identification is based, at least in part, on detection of the unique combination of barcodes.

Provided herein are methods and kits for parallel detection one or more target sequences across multiple samples, comprising separating a set of samples into one or more pooled sets, wherein each sample may comprise an initial amplicon comprising one or more target sequences and at least one barcode; conducting an amplification reaction on the one or more pooled sets to further amplify the amplicons, and optionally further adding an additional barcode to the amplicon; sequencing the amplicons; identifying individual samples from the pooled sample set that are positive for the one or more target sequences based on sequencing of the amplicons, wherein identification is based, at least in part, on detection of the unique combination of barcodes.