Abstract: Provided is a nucleic acid construct for expressing a gene of interest, the nucleic acid construct comprising, in order from 5?-terminus, each sequence of: (a) a 5? long terminal repeat (LTR) sequence derived from a retrovirus; (b) a packaging signal sequence (IV) derived from a retrovirus; (c) a sequence or multiple cloning site of the gene of interest; (d) a post-transcriptional regulatory sequence (PRE); (e) an siRNA generating sequence which forms at least one stem-loop structure and in which RNA, which induces RNA interference in mammalian cells, is transcribed; and (f) a 3? LTR sequence derived form a retrovirus.

Abstract: The present invention regards a variety of methods and compositions for whole genome amplification and whole transcriptome amplification. In a particular aspect of the present invention, there is a method of amplifying a genome comprising a library generation step followed by a library amplification step. In specific embodiments, the library generating step utilizes specific primer mixtures and a DNA polymerase, wherein the specific primer mixtures are designed to eliminate ability to self-hybridize and/or hybridize to other primers within a mixture but efficiently and frequently prime nucleic acid templates.

Abstract: Methods of producing an amplified double stranded deoxyribonucleic acid (dsDNA) from a nucleic acid sample are provided. Aspects of the methods include amplifying using a single product nucleic acid primer and a template switch oligonucleotide to produce an amplified dsDNA product. Compositions and kits for use in performing the methods are also provided.

Abstract: The purpose of the present invention is to efficiently produce microglia from pluripotent stem cells. Provided is a method for producing microglia from pluripotent stem cells, comprising the following steps: (a) a step of co-culturing a pluripotent stem cell together with a feeder cell for 7 days or longer, and obtaining a blood progenitor cell; (b) a step of co-culturing the blood progenitor cell obtained in step (a) together with a feeder cell in the presence of IL-3 and/or GM-CSF, and obtaining an embryonic monocyte; and (c) a step of, in the presence of M-CSF, co-culturing the embryonic monocyte obtained in step (b) together with an astrocyte, or culturing the embryonic monocyte using an astrocyte supernatant.

Abstract: Provided are methods, devices, assemblies, and systems for dispensing into the wells of a multi-well device and imaging such wells from multiple Z-planes. Multi-Z imaging of the present methods and systems may allow for the detection of wells of a multi-well device that contain a desired number of cells. Also provided are methods, devices, assemblies, and systems for processing cell-containing wells of a multi-well device identified through the use of multi-Z imaging.

Abstract: A method for producing a nucleic acid-encapsulated adeno-associated virus (AAV) hollow particle, comprising the following steps: (1) preparing a linear nucleic acid fragment comprising a sequence for A region and a sequence for D? region in an AAV inverted terminal repeat (ITR) (i.e., an AD sequence) or a sequence complementary to the AD sequence and a target gene sequence; (2) introducing the nucleic acid fragment prepared in step (1) into a cell capable of producing an AAV hollow particle; and (3) culturing the cell obtained in step (2).

Abstract: The present disclosure provides methods, compositions, and systems employing blocked primers. Aspects of the disclosure include providing a blocked primer reaction mixture that includes a blocked primer and a template nucleic acid component from a single cell; unblocking the blocked primer to produce an active primer reaction mixture and subjecting the activated primer reaction mixture to primer extension conditions, such as nucleic acid amplification conditions.

Abstract: The present invention provides a nucleic acid which encodes an adeno-associated virus (AAV) capsid protein mutant that contains a peptide comprising an amino acid sequence selected from the group consisting of SEQ ID Nos. 15 to 62 or a peptide comprising an amino acid sequence produced by substituting, deleting, inserting and/or adding one or several amino acid residues in an amino acid sequence selected from the group consisting of SEQ ID Nos. 15 to 62; DNA comprising the nucleic acid; a cell harboring the DNA; and a method for producing the cell.

Abstract: Provided is a green tea-containing beverage including: (A) green tea; (B) at least one selected from the group consisting of phytic acid, tartaric acid, and gluconic acid; and (C) at least one selected from the group consisting of ascorbic acid, a salt of ascorbic acid, erythorbic acid, and a salt of erythorbic acid, wherein the green tea-containing beverage has a copper content of 0.3 mg/L or more. Preferably, the concentration of the constituent (B) is 0.05-2.5 g/L. Preferably, the concentration of the constituent (C) is 0.2-5 g/L. Preferably the green tea-containing beverage further includes alcohol. Also provided is a method for preventing discoloration of a green tea-containing beverage, the method including: incorporating the constituents (B) and (C) in the green tea-containing beverage; and further incorporating copper so as to give the copper content of 0.3 mg/L or more.

Abstract: The present invention relates to a method for producing endothelial cells, including carrying out: (a) inducing a population of mesoderm-lineage cells containing endothelial progenitor cells from pluripotent stem cells without forming an embryoid body; and (b) culturing the population of mesoderm-lineage cells containing endothelial progenitor cells in the presence of RepSox, in this order. According to the present invention, endothelial cells with high quality can be efficiently produced from pluripotent stem cells. The endothelial cells obtained by the method of the present invention are useful for the production of, for example, a myocardial sheet, and expected to be utilized in the treatment of a heart disease. A myocardial sheet can be produced by mixing the endothelial cells obtained by the method of the present invention with myocardial cells and mural cells and culturing the cells.

Abstract: Provided are: a reverse transcriptase mutant including an amino acid mutation at a position corresponding to position 55 of the amino acid sequence of wild-type reverse transcriptase derived from the Moloney murine leukemia virus, wherein the reverse transcriptase mutant is characterized in that the amino acid mutation is a substitution from threonine to another amino acid, and the other amino acid is selected from the group consisting of amino acids having a nonpolar aliphatic side chain and amino acids having a polar acidic functional group side chain; a nucleic acid that encodes the mutant; a method for producing the mutant and the nucleic acid that encodes the mutant; a method for synthesizing cDNA in which the mutant is used; and a composition and kit including the mutant.

Abstract: Provided are methods of depleting a target nucleic acid in a sample. The methods include contacting a target nucleic acid with two or more polymers that specifically hybridize to the target nucleic acid, and cleaving the hybridized regions of the target nucleic acid to deplete the target nucleic acid in the sample. Kits for practicing the subject methods are also provided.

Abstract: The present disclosure provides methods, device, assemblies, and systems for dispensing and visualizing single cells. For example, provided herein are systems and methods for dispensing a dispense volume into a plurality of wells of a multi-well device, where, on average, a pre-determined number of cells (e.g., 1-20) are present in the dispense volume, and determining, via a cellular label, the number of cells present in each of the plurality of wells. Such dispensing and cell detection may be repeated a number of times with respect to wells identified as having less than the pre-determined number of cells in order increase the number wells in the multi-well device containing the desired number (e.g., a single cell).

Abstract: Provided are methods of adding adapters to nucleic acids. The methods include combining in a reaction mixture a template nucleic acid, a template switch oligonucleotide, a polymerase, and dNTPs. The reaction mixture components are combined under conditions sufficient to produce a product nucleic acid that includes the template nucleic acid and the template switch oligonucleotide each hybridized to adjacent regions of a single product nucleic acid including a region polymerized from the dNTPs by the polymerase. The methods further include attaching sequencing platform adapter constructs to ends of the product nucleic acid or a derivative thereof. Aspects of the invention further include compositions and kits.

Abstract: Provided are: a reverse transcriptase mutant including an amino acid mutation at a position corresponding to position 55 of the amino acid sequence of wild-type reverse transcriptase derived from the Moloney murine leukemia virus, wherein the reverse transcriptase mutant is characterized in that the amino acid mutation is a substitution from threonine to another amino acid, and the other amino acid is selected from the group consisting of amino acids having a nonpolar aliphatic side chain and amino acids having a polar acidic functional group side chain; a nucleic acid that encodes the mutant; a method for producing the mutant and the nucleic acid that encodes the mutant; a method for synthesizing cDNA in which the mutant is used; and a composition and kit including the mutant.

Abstract: Spin columns that include a poly(acid) membrane separation matrix are provided. Also provided are kits that include the subject devices, as well as methods of using the devices, e.g., in sample preparation (such as protein purification) protocols.

Abstract: Stem cells can be efficiently proliferated by culturing the stem cells in the presence of a novel recombinant fibronectin fragment.

Abstract: The present invention provides: a mutant of adeno-associated virus (AAV) capsid protein, which contains at least one amino acid substitution in PLA2 domain when compared with the amino acid sequence for wild-type AAV capsid protein; a nucleic acid encoding the mutant; a cell containing the nucleic acid; a method for producing a recombinant AAV particle, comprising a step of culturing the cell to produce the recombinant AAV particle; a recombinant AAV particle containing the mutant; a composition containing the recombinant AAV particle; and a method for transferring a gene into a target cell, comprising a step of bringing the recombinant AAV particle into contact with the target cell.

Abstract: The present invention concerns preparation of DNA molecules, such as a library, using a stem-loop oligonucleotide. In particular embodiments, the invention employs a single reaction mixture and conditions. In particular, at least part of the inverted palindrome is removed during the preparation of the molecules to facilitate amplification of the molecules. Thus, in specific embodiments, the DNA molecules are suitable for amplification and are not hindered by the presence of the palindrome.

Abstract: Provided are: a DNA polymerase mutant having reverse transcriptase activity, the DNA polymerase mutant including a sequence consisting of twelve specific amino acids A1-A12, wherein the DNA polymerase mutant having reverse transcriptase activity is characterized in that the A3 and/or A10 amino acid is substituted by a basic amino acid residue different from that prior to the introduction of mutation; a kit and a composition including the DNA polymerase; a method for producing the DNA polymerase; and a method for modifying an existing DNA polymerase having reverse transcriptase activity.