Abstract: The present invention relates to a method, a device, and an apparatus for analyzing the expression of a gene in single cells. Specifically, the present invention relates to: a device for gene expression analysis, characterized by including a support, in which a nucleic acid probe having a test nucleic acid capture sequence and a known sequence, and further containing a cell recognition tag sequence which differs depending on the difference in position on the surface of the support or in the vicinity of the surface thereof, and a common primer sequence having a known sequence is two-dimensionally distributed and immobilized on the surface of the support or in the vicinity of the surface thereof; and a method and an apparatus using the device for gene expression analysis.

Abstract: The present invention provides a method and/or means for collecting and analyzing an individual cell in a tissue, and at the same time, quantitatively monitoring the expression levels of various genes while keeping two-dimensional information in the tissue. Specifically, the present invention provides a method comprising preparing a cDNA library from mRNA while keeping two-dimensional cellular distribution information and obtaining the gene expression levels at any site or all sites at a level of single cell. More specifically, the present invention provides a method comprising preparing a cDNA library in a sheet-form from mRNA while keeping two-dimensional cellular distribution information and repeatedly using the cDNA library in the detection of the gene expression, thereby allowing measurement of the expression distribution for a number of genes at a high accuracy.

Abstract: The present invention relates to a method, a device, and an apparatus for analyzing the expression of a gene in single cells. Specifically, the present invention relates to: a device for gene expression analysis, characterized by including a support, in which a nucleic acid probe having a test nucleic acid capture sequence and a known sequence, and further containing a cell recognition tag sequence which differs depending on the difference in position on the surface of the support or in the vicinity of the surface thereof, and a common primer sequence having a known sequence is two-dimensionally distributed and immobilized on the surface of the support or in the vicinity of the surface thereof; and a method and an apparatus using the device for gene expression analysis.

Abstract: The present invention relates to a method, a device, and an apparatus for analyzing the expression of a gene in single cells. Specifically, the present invention relates to: a device for gene expression analysis, characterized by including a support, in which a nucleic acid probe having a test nucleic acid capture sequence and a known sequence, and further containing a cell recognition tag sequence which differs depending on the difference in position on the surface of the support or in the vicinity of the surface thereof, and a common primer sequence having a known sequence is two-dimensionally distributed and immobilized on the surface of the support or in the vicinity of the surface thereof; and a method and an apparatus using the device for gene expression analysis.

Abstract: A method for analyzing gene expression in a cell using a device, the method including: introducing a plurality of cells into microreactors so that a single cell corresponds to each microreactor, capturing mRNA from the single cell on the probe, synthesizing first cDNA by subjecting the captured mRNA to a reverse transcription reaction, to produce a first cDNA library derived from the single cell on the solid supports, washing the solid supports, a step of synthesizing second cDNA from the first cDNA library, performing fragmentation of double-stranded DNA containing the first and second cDNA and addition of a tag sequence, removing a component other than an immobilized double-stranded DNA fragment by washing the solid supports with a washing solution, performing amplification of the double-stranded DNA fragment, and performing, for the amplified sequence, analysis of gene expression in each single cell, using the cell identification sequence and the molecule identification sequence.

Abstract: In order to conduct gene expression analysis of a number of genes in a number of cells, it has been necessary to separate cells, extract genes therefrom, amplify nucleic acids, and perform sequence analysis. However, separation of cells imposes damages on the cells, and it requires the use of an expensive system. Gene expression analysis in each cell can be carried out with high accuracy by arranging a pair of structures comprising a cell trapping section and a nucleic acid trapping section in a vertical direction to extract individual genes in relevant cells, synthesizing cDNA in the nucleic acid trapping section, amplifying nucleic acids, and analyzing the sequences using a next-generation sequencer.

Abstract: The present invention provides a method and/or means for collecting and analyzing an individual cell in a tissue, and at the same time, quantitatively monitoring the expression levels of various genes while keeping two-dimensional information in the tissue. Specifically, the present invention provides a method comprising preparing a cDNA library from mRNA while keeping two-dimensional cellular distribution information and obtaining the gene expression levels at any site or all sites at a level of single cell. More specifically, the present invention provides a method comprising preparing a cDNA library in a sheet-form from mRNA while keeping two-dimensional cellular distribution information and repeatedly using the cDNA library in the detection of the gene expression, thereby allowing measurement of the expression distribution for a number of genes at a high accuracy.

Abstract: In order to conduct gene expression analysis of a number of genes in a number of cells, it has been necessary to separate cells, extract genes therefrom, amplify nucleic acids, and perform sequence analysis. However, separation of cells imposes damages on the cells, and it requires the use of an expensive system. Gene expression analysis in each cell can be carried out with high accuracy by arranging a pair of structures comprising a cell trapping section and a nucleic acid trapping section in a vertical direction to extract individual genes in relevant cells, synthesizing cDNA in the nucleic acid trapping section, amplifying nucleic acids, and analyzing the sequences using a next-generation sequencer.

Abstract: The present invention aims to provide a data analysis apparatus capable of clustering appropriately even when there is an exceptional datum resulted from an experimental error and the like. In the data analysis apparatus according to the invention, a cluster range parameter for stretching a cluster boundary is determined in advance according to the range of an experimental error which an experimental error datum describes. In the process of clustering, an exceptional datum which does not belong to any cluster is determined to belong to a cluster when an area at a distance determined by the cluster range parameter from the exceptional datum is contained in the cluster, and the exceptional datum is determined to form an independent cluster when even the area at the distance is not contained in any cluster (see FIG. 7).

Abstract: According to a semiconductor device (101), a first switching active element (103) of a normally-on type and a second switching active element (104) of a normally-off type are cascode-connected to each other. This causes an electric current path to be formed. The first and second switching active elements (103, 104) are provided and connected so that loop area of the electric current path is a minimum area in a plan view of the semiconductor device (101).

Abstract: The present invention relates to a method, a device, and an apparatus for analyzing the expression of a gene in single cells. Specifically, the present invention relates to: a device for gene expression analysis, characterized by including a support, in which a nucleic acid probe having a test nucleic acid capture sequence and a known sequence, and further containing a cell recognition tag sequence which differs depending on the difference in position on the surface of the support or in the vicinity of the surface thereof, and a common primer sequence having a known sequence is two-dimensionally distributed and immobilized on the surface of the support or in the vicinity of the surface thereof; and a method and an apparatus using the device for gene expression analysis.

Abstract: A method of synthesizing cDNA, in sequence, includes picking up a whole single-cell from a sample containing at least a single-cell having a cell membrane, lysing the cell membrane and extracting nucleic acids from the cell, degrading DNA of the extracted nucleic acids with DNase, hybridizing mRNA of the total RNA contained in the whole single-cell with oligo (dT) fixed onto a carrier, performing reverse transcription of the mRNA hybridized with the oligo (dT) to fix cDNA derived from the single-cell onto the carrier, thereby preparing a single-cell derived cDNA library fixed onto a carrier, and amplifying cDNA fixed onto the carrier.

Abstract: According to a semiconductor device (101), a first switching active element (103) of a normally-on type and a second switching active element (104) of a normally-off type are cascode-connected to each other. This causes an electric current path to be formed. The first and second switching active elements (103, 104) are provided and connected so that loop area of the electric current path is a minimum area in a plan view of the semiconductor device (101).

Abstract: It is an object to provide a method of suitably analyzing the amount of gene expression of a single-cell. A method of detecting a nucleic acid comprising a step of sampling a single-cell from a sample containing at least a single-cell, a cell lysis step of lysing cell membrane of the sampled single-cell and extracting nucleic acids from the cell, a DNase treatment step of degrading DNA of the extracted nucleic acids with DNase, a step of hybridizing mRNA of the total RNA contained in the single-cell with oligo (dT) fixed onto a carrier, a step of performing reverse transcription of the mRNA hybridized with the oligo (dT) to fix cDNA derived from the single-cell onto the carrier, thereby preparing a single-cell derived cDNA library fixed onto a carrier, and a step of amplifying cDNA fixed onto the carrier and simultaneously detecting an amplification amount of the cDNA.

Abstract: It is intended to provide a novel convenient approach for DNA quantitative analysis that overcomes the disadvantages of conventional formulations. A standard DNA sample is prepared by introducing a single-base substitution into target DNA, and a predetermined amount thereof is mixed with a target DNA sample. The target and standard DNAs are amplified using the same primers designed to amplify a region comprising the single-base substitution site. To a hybridization product of a probe capable of binding to a site immediately before the single-base substitution site, ddATP, ddGTP, ddCTP, and ddTTP are sequentially added one by one to perform a complementary strand synthesis reaction. Luciferase reaction-induced luminescence derived from the formed pyrophosphoric acid is detected. The target DNA is quantitated from the amount of the detected luminescence and the amount of the added standard DNA sample.

Abstract: The present invention provides a method and/or means for collecting and analyzing an individual cell in a tissue, and at the same time, quantitatively monitoring the expression levels of various genes while keeping two-dimensional information in the tissue. Specifically, the present invention provides a method comprising preparing a cDNA library from mRNA while keeping two-dimensional cellular distribution information and obtaining the gene expression levels at any site or all sites at a level of single cell. More specifically, the present invention provides a method comprising preparing a cDNA library in a sheet-form from mRNA while keeping two-dimensional cellular distribution information and repeatedly using the cDNA library in the detection of the gene expression, thereby allowing measurement of the expression distribution for a number of genes at a high accuracy.

Abstract: It is intended to provide a novel convenient approach for DNA quantitative analysis that overcomes the disadvantages of conventional formulations. A standard DNA sample is prepared by introducing a single-base substitution into target DNA, and a predetermined amount thereof is mixed with a target DNA sample. The target and standard DNAs are amplified using the same primers designed to amplify a region comprising the single-base substitution site. To a hybridization product of a probe capable of binding to a site immediately before the single-base substitution site, ddATP, ddGTP, ddCTP, and ddTTP are sequentially added one by one to perform a complementary strand synthesis reaction. Luciferase reaction-induced luminescence derived from the formed pyrophosphoric acid is detected. The target DNA is quantitated from the amount of the detected luminescence and the amount of the added standard DNA sample.

Abstract: It is an object to provide a method of suitably analyzing the amount of gene expression of a single-cell. A method of detecting a nucleic acid comprising a step of sampling a single-cell from a sample containing at least a single-cell, a cell lysis step of lysing cell membrane of the sampled single-cell and extracting nucleic acids from the cell, a DNase treatment step of degrading DNA of the extracted nucleic acids with DNase, a step of hybridizing mRNA of the total RNA contained in the single-cell with oligo (dT) fixed onto a carrier, a step of performing reverse transcription of the mRNA hybridized with the oligo (dT) to fix cDNA derived from the single-cell onto the carrier, thereby preparing a single-cell derived cDNA library fixed onto a carrier, and a step of amplifying cDNA fixed onto the carrier and simultaneously detecting an amplification amount of the cDNA.

Abstract: A simple and highly accurate method for detecting the presence or absence of gene mutation and methylated cytosine in CpG dinucleotide that are contained in a target sequence derived from an analysis sample is provided. Features of the method for nucleic acid analysis include cleaving one or more noncomplementary sites in a double-stranded nucleic acid sample by a single strand-specific endonuclease, hybridizing at least one of the nucleic acid fragments obtained to a probe containing a nucleotide sequence that is partially or totally identical to either one strand of the double-stranded nucleic acid sample, allowing an extension reaction to proceed from the nucleic acid fragment hybridized to the probe, and optically detecting pyrophosphate generated by the extension reaction, thereby judging the presence or absence of at least a noncomplementary site in the double-stranded nucleic acid sample.

Abstract: This invention provides a method of genetic testing that enables testing of a plurality of variation sites (SNPs) in a cost-effective and simple manner, allowing realization of genetic diagnosis in clinical settings. The SNP type of the nucleic acid sample is evaluated by: allowing a nucleic acid sample having an anchor sequence at its 5? end to hybridize to a support having, immobilized on its surface, a probe containing a sequence that is complementary to the target sequence (the SNP region); extending a complementary strand from the probe utilizing the nucleic acid sample as a template; dissociating and removing the nucleic acid sample from the extended probe; extending a complementary strand using the extended probe as a template and a primer having a sequence identical to the anchor sequence; and detecting pyrophosphoric acid generated via the primer extension, based on bioluminescence.