The present invention relates to a method and a kit for detecting alleles of which the specificity and sensitivity in a DNA polymerase chain reaction (PCR), which is widely used for the detection of minor alleles such as single nucleotide polymorphisms or somatic mutations, are increased. More specifically, the present invention relates to a PCR-based method and kit for single nucleotide polymorphism (SNP) genotyping and somatic mutation detection, the method and kit adding a partially or fully double-stranded oligonucleotide for increasing discrimination to a PCR solution for selective amplification of alleles, so that PCR amplification is not affected when a primer 3′ terminal base is complementary (3′-matched) to a template but PCR amplification is strongly inhibited when a 3′ terminal base is not complementary (3′-mismatched).

The present invention relates to a method and a kit for detecting alleles of which the specificity and sensitivity in a DNA polymerase chain reaction (PCR), which is widely used for the detection of minor alleles such as single nucleotide polymorphisms or somatic mutations, are increased. More specifically, the present invention relates to a PCR-based method and kit for single nucleotide polymorphism (SNP) genotyping and somatic mutation detection, the method and kit adding a partially or fully double-stranded oligonucleotide for increasing discrimination to a PCR solution for selective amplification of alleles, so that PCR amplification is not affected when a primer 3′ terminal base is complementary (3′-matched) to a template but PCR amplification is strongly inhibited when a 3′ terminal base is not complementary (3′-mismatched).

Provided are devices, systems, and methods for the identification, quantification, and profiling of microscopic organisms. The methods for the identification, quantification, and profiling of microscopic organisms include, for example, the selective enrichment of microscopic organisms from a heterogeneous sample; subsequent loading of the microscopic organisms into microfluidic channels or reaction chambers; direct amplification of nucleic acids from single, isolated microscopic organisms; and examination of amplification products using digital High Resolution Melting (HRM) analysis.

Provided are devices, systems, and methods for the identification, quantification, and profiling of microscopic organisms. The methods for the identification, quantification, and profiling of microscopic organisms include, for example, the selective enrichment of microscopic organisms from a heterogeneous sample; subsequent loading of the microscopic organisms into microfluidic channels or reaction chambers; direct amplification of nucleic acids from single, isolated microscopic organisms; and examination of amplification products using digital High Resolution Melting (HRM) analysis.

The present invention relates to a method for identifying a microorganism in a biological sample by polymerase chain reaction (PCR), comprising the steps of a) providing a biological sample suspected of comprising microbes, and optionally isolating nucleic acid sequences from said biological sample; b) PCR amplifying at least one microbial rRNA internal transcribed spacer (ITS) region comprised in said optionally isolated nucleic acid sequences using a set of broad-taxonomic range amplification primers to thereby generate PCR amplicons from nucleic acid sequences of microbial origin; c) recording a high resolution melting curve for the PCR amplicons, and recording the length of the PCR amplicons; d) comparing the high resolution melting curve with a database comprising high resolution melting curves of reference amplicons of known microbial species or strains, to thereby obtain a first identity indicator; e) comparing the length of each PCR amplicon having a distinct length with a database comprising PCR amplicon lengths of reference amplicons of known microbial species or strains, to thereby obtain a second identity indicator; and f) identifying the microorganism present in said sample to the species or strain level if the first and second identity indicator match.

The present invention relates to a method for identifying a microorganism in a biological sample by polymerase chain reaction (PCR), comprising the steps of a) providing a biological sample suspected of comprising microbes, and optionally isolating nucleic acid sequences from said biological sample; b) PCR amplifying at least one microbial rRNA internal transcribed spacer (ITS) region comprised in said optionally isolated nucleic acid sequences using a set of broad-taxonomic range amplification primers to thereby generate PCR amplicons from nucleic acid sequences of microbial origin; c) recording a high resolution melting curve for the PCR amplicons, and recording the length of the PCR amplicons; d) comparing the high resolution melting curve with a database comprising high resolution melting curves of reference amplicons of known microbial species or strains, to thereby obtain a first identity indicator; e) comparing the length of each PCR amplicon having a distinct length with a database comprising PCR amplicon lengths of reference amplicons of known microbial species or strains, to thereby obtain a second identity indicator; and f) identifying the microorganism present in said sample to the species or strain level if the first and second identity indicator match.

Compositions, kits, and methods for performing rapid polymerase chain reaction (PCR) to amplify a target nucleic acid in a biological sample are disclosed. The methods include the use of at least one hybridization stabilizer and/or the adjustment of the thermocycling profiles between initiation and propagation phases of the amplification process. Also disclosed are methods of detecting the target nucleic acid following amplification thereof, as well as reaction mixtures that may be utilized in said methods.

Compositions, kits, and methods for performing rapid polymerase chain reaction (PCR) to amplify a target nucleic acid in a biological sample are disclosed. The methods include the use of at least one hybridization stabilizer and/or the adjustment of the thermocycling profiles between initiation and propagation phases of the amplification process. Also disclosed are methods of detecting the target nucleic acid following amplification thereof, as well as reaction mixtures that may be utilized in said methods.

Provided is a method for detecting and quantifying a nucleic acid in real time and at high speed. The present disclosure provides a real-time high-speed PCR method in which fluorescent signals can be measured from a single-wavelength light source by using a single signal fluorescent material under continuous temperature control. Thus, the PCR method can be performed with a compact lightweight device with a simplified structure.

Provided is a method for detecting and quantifying a nucleic acid in real time and at high speed. The present disclosure provides a real-time high-speed PCR method in which fluorescent signals can be measured from a single-wavelength light source by using a single signal fluorescent material under continuous temperature control. Thus, the PCR method can be performed with a compact lightweight device with a simplified structure.