Methods and systems are provided for massively parallel genetic analysis of single cells in emulsion droplets or reaction containers. Genetic loci of interest are targeted in a single cell using a set of probes, and a fusion complex is formed by molecular linkage and amplification techniques. Methods are provided for high-throughput, massively parallel analysis of the fusion complex in a single cell in a population of at least 10,000 cells. Also provided are methods for tracing genetic information back to a cell using barcode sequences.

Methods and systems are provided for massively parallel genetic analysis of single cells in emulsion droplets or reaction containers. Genetic loci of interest are targeted in a single cell using a set of probes, and a fusion complex is formed by molecular linkage and amplification techniques. Methods are provided for high-throughput, massively parallel analysis of the fusion complex in a single cell in a population of at least 10,000 cells. Also provided are methods for tracing genetic information back to a cell using barcode sequences.

Colorimetry is used to detect amplification reaction products. A sample is contacted with a reaction mix under conditions such that an amplification reaction occurs and produces an amplification reaction product if the sample contains a target nucleic acid template molecule. The reaction mix includes an enzyme for catalyzing the amplification reaction, and at least one halochromic agent. If the target nucleic acid template molecule is present, the amplification reaction changes the starting pH of the reaction mix to cause a detectable colorimetric change of the halochromic agent, thereby indicating the presence of the target nucleic acid. If the target nucleic acid template molecule is not present, the amplification reaction does not generate an adequate number of protons to sufficiently change the starting pH of the reaction mix to cause a detectable colorimetric change of the halochromic agent, thereby indicating that the amplification reaction product has not been produced.

Colorimetry is used to detect amplification reaction products. A sample is contacted with a reaction mix under conditions such that an amplification reaction occurs and produces an amplification reaction product if the sample contains a target nucleic acid template molecule. The reaction mix includes an enzyme for catalyzing the amplification reaction, and at least one halochromic agent. If the target nucleic acid template molecule is present, the amplification reaction changes the starting pH of the reaction mix to cause a detectable colorimetric change of the halochromic agent, thereby indicating the presence of the target nucleic acid. If the target nucleic acid template molecule is not present, the amplification reaction does not generate an adequate number of protons to sufficiently change the starting pH of the reaction mix to cause a detectable colorimetric change of the halochromic agent, thereby indicating that the amplification reaction product has not been produced.

Systems and methods for detection of a signal from droplets of an emulsion. An exemplary system may comprise a fluid transporter having a tube with an open end for aspirating droplets, a singulator to arrange the droplets in single file and to space the single-file droplets from one another, and a detection channel in optical communication with a detector configured to detect a signal from droplets. In some embodiments, the singulator may have a channel junction at which a stream of droplets in single file is combined with a stream of spacing fluid, and a tapered spacing channel extending downstream from the channel junction toward the detection channel. In some embodiments, the fluid transporter may suck droplet-containing fluid and spacing fluid through the detection channel from respective sources. In some embodiments, droplets may be subjected to a disaggregation routine before they are passed through the detection channel.

A detection method including bringing liquid including a target nucleic acid into contact with a well array having wells such that at most one molecule of the target nucleic acid is included per well, sealing the well such that the target nucleic acid remains in the well, amplifying, in the well, a signal derived from the target nucleic acid, detecting the signal emitted from the well, and detecting whether the target nucleic acid includes at least one of a first nucleic acid, a second nucleic acid, and a double-stranded nucleic acid resulting from complementary binding of the first and second nucleic acids. The signal includes at least one of a first signal emitted by binding of a first specific binding substance to the first nucleic acid, and a second signal different from the first signal and emitted by binding of a second specific binding substance to the second nucleic acid.

A detection method including bringing liquid including a target nucleic acid into contact with a well array having wells such that at most one molecule of the target nucleic acid is included per well, sealing the well such that the target nucleic acid remains in the well, amplifying, in the well, a signal derived from the target nucleic acid, detecting the signal emitted from the well, and detecting whether the target nucleic acid includes at least one of a first nucleic acid, a second nucleic acid, and a double-stranded nucleic acid resulting from complementary binding of the first and second nucleic acids. The signal includes at least one of a first signal emitted by binding of a first specific binding substance to the first nucleic acid, and a second signal different from the first signal and emitted by binding of a second specific binding substance to the second nucleic acid.

An object of the present invention is to provide a new freeze-dried article that is easy to handle and a producing method thereof. A freeze-dried structure of the present invention includes a freeze-dried article and a support member, wherein the support member has an embedded region and a protruding region, the embedded region of the support member is embedded in the internal portion of the freeze-dried article, and the protruding region of the support member protrudes outward from the freeze-dried article.

An object of the present invention is to provide a new freeze-dried article that is easy to handle and a producing method thereof. A freeze-dried structure of the present invention includes a freeze-dried article and a support member, wherein the support member has an embedded region and a protruding region, the embedded region of the support member is embedded in the internal portion of the freeze-dried article, and the protruding region of the support member protrudes outward from the freeze-dried article.

The present invention relates to an apparatus for performing a nucleic acid amplification reaction and a fluorescence detection device for reaction analysis. The nucleic acid amplification apparatus of the present invention uses a plurality of blocks having different reaction temperatures by independent temperature control and the movement between the blocks is performed along sliding recesses formed in the blocks, enabling to greatly shorten the total amplification time (TAT). In the fluorescence detection device of the present invention, the positions of the light source and the photodetector are very unique for the reaction vessel in which an excitation light is provided and an emission light is generated.