A nucleic acid amplification reagent includes a probe which anneals to a target nucleic acid contained in a nucleic acid, and an intercalator which is inserted between base pairs of one strand of the nucleic acid and a complementary strand synthesized on the one strand, and between base pairs of the other strand of the nucleic acid and a complementary strand synthesized on the other strand. The wavelength band of light emitted from the probe and the wavelength band of light emitted from the intercalator at least partially overlap with each other.

A nucleic acid amplification reagent includes a probe which anneals to a target nucleic acid contained in a nucleic acid, and an intercalator which is inserted between base pairs of one strand of the nucleic acid and a complementary strand synthesized on the one strand, and between base pairs of the other strand of the nucleic acid and a complementary strand synthesized on the other strand. The wavelength band of light emitted from the probe and the wavelength band of light emitted from the intercalator at least partially overlap with each other.

An object of the invention is to provide a primer for detecting and quantifying homozygous deletion of the SMN1 gene, the deletion of which causes SMA, using a dried blood spot in a filter paper and a method related to specific detection/quantification of the SMN1 gene using the primer. The invention is a method for detecting the SMN1 gene in a dried blood spot in a filter paper by real-time PCR, including the steps of (A) to (D) below: (A) a step of adding the dried blood spot in a filter paper to a PCR reaction tube; (B) a step of adding a PCR reagent to the PCR reaction tube, wherein the PCR reagent contains at least a primer designed in a manner that the reactivity to the SMN2 gene is less than 1% of that to the SMN1 gene, a polymerase, dNTPs and an intercalator or a fluorescently labeled probe; (C) a step of performing PCR reaction in the tube containing the PCR reagent and the dried blood spot in a filter paper; and (D) a step of sequentially and optically detecting a target nucleic acid in the SMN1 gene amplified by the PCR reaction.

An object of the invention is to provide a primer for detecting and quantifying homozygous deletion of the SMN1 gene, the deletion of which causes SMA, using a dried blood spot in a filter paper and a method related to specific detection/quantification of the SMN1 gene using the primer. The invention is a method for detecting the SMN1 gene in a dried blood spot in a filter paper by real-time PCR, including the steps of (A) to (D) below: (A) a step of adding the dried blood spot in a filter paper to a PCR reaction tube; (B) a step of adding a PCR reagent to the PCR reaction tube, wherein the PCR reagent contains at least a primer designed in a manner that the reactivity to the SMN2 gene is less than 1% of that to the SMN1 gene, a polymerase, dNTPs and an intercalator or a fluorescently labeled probe; (C) a step of performing PCR reaction in the tube containing the PCR reagent and the dried blood spot in a filter paper; and (D) a step of sequentially and optically detecting a target nucleic acid in the SMN1 gene amplified by the PCR reaction.

Compositions, kits, and methods are provided for the normalization of a quantitative polymerase chain reaction (PCR) amplification. Also provided are compositions, kits, and methods for multiplexing qPCR amplification of two or more target nucleic acids in the same well.

Compositions, kits, and methods are provided for the normalization of a quantitative polymerase chain reaction (PCR) amplification. Also provided are compositions, kits, and methods for multiplexing qPCR amplification of two or more target nucleic acids in the same well.

Disclosed herein is a method for the detection of the presence of sperm DNA fragmentation in a semen sample. The method comprises a step of embedding the semen sample containing sperm cells in a gel comprising acrylamide, acrylic acid, methacrylic acid, N-isopropylacrylamide (NIPAM), alginate, or polyethylene glycol (PEG), to obtain a sperm cells-embedded gel. A kit for detecting sperm DNA fragmentation in a semen sample is also disclosed.

Disclosed herein is a method for the detection of the presence of sperm DNA fragmentation in a semen sample. The method comprises a step of embedding the semen sample containing sperm cells in a gel comprising acrylamide, acrylic acid, methacrylic acid, N-isopropylacrylamide (NIPAM), alginate, or polyethylene glycol (PEG), to obtain a sperm cells-embedded gel. A kit for detecting sperm DNA fragmentation in a semen sample is also disclosed.

The present invention provides a fluorogenic RNA capable of visualizing mRNA in cells, particularly, living mammalian cells, and a target RNA fluorescent labeling method using the fluorogenic RNA. The present invention provides a fluorogenic nucleic acid molecule containing a base sequence having two or more fluorescent molecule-binding regions linked via a linker sequence, in which one or more fluorescent molecule-binding aptamer sequences are inserted into a scaffold sequence of each of the fluorescent molecule-binding regions.

The present invention provides a fluorogenic RNA capable of visualizing mRNA in cells, particularly, living mammalian cells, and a target RNA fluorescent labeling method using the fluorogenic RNA. The present invention provides a fluorogenic nucleic acid molecule containing a base sequence having two or more fluorescent molecule-binding regions linked via a linker sequence, in which one or more fluorescent molecule-binding aptamer sequences are inserted into a scaffold sequence of each of the fluorescent molecule-binding regions.