The present disclosure is directed to compositions, methods and kits for amplifying target nucleic acids while reducing non-specific amplification and undesired amplification products using a dual hot start reaction mixture that comprise at least two different hot start mechanisms.

The present disclosure is directed to compositions, methods and kits for amplifying target nucleic acids while reducing non-specific amplification and undesired amplification products using a dual hot start reaction mixture that comprise at least two different hot start mechanisms.

A microfluidic device is described. The device comprises a reaction chamber, wherein the reaction chamber comprises at least one reaction reagent disposed on at least one inner surface of the reaction chamber. A heater is also provided. A thermally dissolvable or degradable or thermally degradable film is applied to the at least one inner surface of the reaction chamber on which the reaction reagent is disposed. Also described is a PCR apparatus and a method of performing PCR.

A microfluidic device is described. The device comprises a reaction chamber, wherein the reaction chamber comprises at least one reaction reagent disposed on at least one inner surface of the reaction chamber. A heater is also provided. A thermally dissolvable or degradable or thermally degradable film is applied to the at least one inner surface of the reaction chamber on which the reaction reagent is disposed. Also described is a PCR apparatus and a method of performing PCR.

A composition is disclosed for the amplification of nucleic acid sequences, which is intended to be mixed with a biological sample containing one or more target nucleic acid sequences and with primers specific for the target nucleic acid sequence(s), and fluorescent probes for revealing an amplification of the target nucleic acid sequence(s). The composition includes reagents and enzymes for nucleic acid application. The composition also includes a polymerizable reagent, which polymerizes under the influence of a stimulation chosen from a heat stimulation and a light stimulation, so as to form a hydrogel which locally encapsulates each target nucleic acid sequence when the composition is exposed to the stimulation. A method using said composition is also disclosed.

A composition is disclosed for the amplification of nucleic acid sequences, which is intended to be mixed with a biological sample containing one or more target nucleic acid sequences and with primers specific for the target nucleic acid sequence(s), and fluorescent probes for revealing an amplification of the target nucleic acid sequence(s). The composition includes reagents and enzymes for nucleic acid application. The composition also includes a polymerizable reagent, which polymerizes under the influence of a stimulation chosen from a heat stimulation and a light stimulation, so as to form a hydrogel which locally encapsulates each target nucleic acid sequence when the composition is exposed to the stimulation. A method using said composition is also disclosed.

A method, comprising amplifying a nucleic acid sequence of interest in a sample comprising genomic DNA of a subject; amplifying a reference nucleic acid sequence in the sample; quantifying the amplified sequence of interest relative to the amplified reference sequence; and determining a copy number of the sequence of interest from the relative quantified amplified sequence of interest. The reference sequence may have at least 80% sequence identity to at least one of SEQ ID NO:1-38, such as SEQ ID NO:1-13. Also disclosed are kits and compositions, each comprising a first probe which specifically hybridizes to at least a portion of at least one reference sequence. Also disclosed is a system configured to perform the above method.

A method, comprising amplifying a nucleic acid sequence of interest in a sample comprising genomic DNA of a subject; amplifying a reference nucleic acid sequence in the sample; quantifying the amplified sequence of interest relative to the amplified reference sequence; and determining a copy number of the sequence of interest from the relative quantified amplified sequence of interest. The reference sequence may have at least 80% sequence identity to at least one of SEQ ID NO:1-38, such as SEQ ID NO:1-13. Also disclosed are kits and compositions, each comprising a first probe which specifically hybridizes to at least a portion of at least one reference sequence. Also disclosed is a system configured to perform the above method.

A portable diagnostic device has a lysate stage (167) with a port for receiving a sample and containing magnetic beads with a probe, and an outlet port. A series of assay stages (161-164) are linked with the lysate vessel, each with a reservoir linked by channels. The final stage (164) has a sensor (169) for detecting beads attached to analyte molecules which have been conveyed according to attachment to probes on beads. Larger transport beads cause reporter beads which are tethered by target NA and probes to be transported to the final sensor stage, where they are released and detected when the transport beads have been removed.

A portable diagnostic device has a lysate stage (167) with a port for receiving a sample and containing magnetic beads with a probe, and an outlet port. A series of assay stages (161-164) are linked with the lysate vessel, each with a reservoir linked by channels. The final stage (164) has a sensor (169) for detecting beads attached to analyte molecules which have been conveyed according to attachment to probes on beads. Larger transport beads cause reporter beads which are tethered by target NA and probes to be transported to the final sensor stage, where they are released and detected when the transport beads have been removed.