Abstract: Described herein are techniques for assembling a polynucleotide encoding a transcription activator-like effector nucleases (TALEN). The techniques ligate and digest necessary modules for a TALEN assembly in one reactor or system. Methods and Kits for generating a TALEN are also described.

Abstract: Microfluidic devices of the present disclosure relate to quick and inexpensive microfluidic manipulation/handling. A number of channels may be supplied with fluid ingredient(s). In some embodiments, a number of protrusions as well as a sealing material may be disposed adjacent to the channels. When the channels are supplied with fluid ingredient(s), the channels may be partitioned into a number of separate cavities that are fluidly isolated from one another. For instance, a sealing material may be compressed so as to deform into the channels, obstructing fluid flow. In some embodiments, the channels supply fluid ingredients to a number of pre-formed cavities. Once the cavities are supplied with fluid ingredient, channels connecting the cavities may be sealed off; that is, the cavities may be subject to fluid isolation. When appropriate, contents within reaction chambers may be subject to further processing (e.g., thermal cycling, various analyses).

Abstract: Procedure for the specific isolation of total DNA content of bacterial germs of different samples, in the course of which the cells are lysated, the DNA content of the lysate is bound selectively, it is washed and then the desalinated linear polymer nucleic acid is eluted from the binding surface in an aqueous solution. Before cell lysis the nonviable bacterial cells are separated from the viable cells on the basis of their different cell surface physical-chemical characteristics, the viable cells of the sample are kept and then lysated using a mechanical and/or enzymatic, favorably lysozyme enzymatic method. After this exclusively double-stranded DNA deriving from the lysate of viable cells is bound on a —SiO2—TiO2- matrix containing chemically activated —OH and dodecylamine groups, and after washing it, the desalinated linear polymer nucleic acid is eluted in an aqueous solution.

Abstract: A method of providing protection against pneumococcal infection in a subject is disclosed. The method includes steps of administering to the subject a composition that includes combination of three recombinant pneumococcal neuraminidases: NanA, NanB, and NanC of S. pneumoniae strains CGSP14, wherein administration of the recombinant pneumococcal neuraminidases elicits an immune response to S. pneumoniae, and treats the subject. In one embodiment, the method further includes a step of adding adjuvants to enhance the immune response. The method also includes a step of using passive antibodies, wherein said passive antibodies are anti-neuraminidase antibodies generated from neuraminidases-immunized humanized animals: NanA, NanB, and NanC. Meanwhile, this invention also provides a method for the molecular diagnosis of pneumococcal infection.

Abstract: Systems and methods for preparing a sample for further analysis are provided. The system can include an enclosure. A membrane can be disposed within the enclosure. First and second reservoirs can be disposed within the enclosure, and at least one of the first and second reservoirs can be adapted to have a reagent disposed therein. A valve can be disposed within the enclosure and in fluid communication with the first or second reservoirs or both. The valve can also be in fluid communication with the membrane. The valve can be adapted to selectively regulate the flow of the reagent from the first reservoir, through the membrane, and into the second reservoir.

Abstract: The methods of the present invention provide methods for manufacturing a master substrate and methods for manufacturing replica arrays from the master substrate. The methods may be used, for example, directly to manufacture or “print” peptide arrays from a DNA array; however, the methods are applicable to a wide range of manufacturing applications for use any time multiple copies of an array needs to be printed.

Abstract: A gene chip is provided for detection. When used on clinical detection, the gene chip directly detects blood without magnification. Gene expressions are examined with high sensitivity and convenience simultaneously.

Abstract: The invention provides systems, devices, and methods for heating and cooling chemical or biological samples, such as genetic materials during Polymerase Chain Reaction (“PCR”). The systems, devices, and methods comprise use of a fluid that performs repeated heating and cooling cycles, e.g., ‘thermal cycling’, on sample reactants with a phase changing fluid during evaporation and condensation. The systems, devices, and methods eliminate the need for a heating block as a means to obtain fast and uniform thermal cycling. The disclosure also describes the use of an optical system in conjunction with the thermodynamic cycler for real-time detection. Ultimately, uniformity and speed of the thermodynamic cycler provides for higher sensitivity and throughput of gene replication and detection.

Abstract: A method for analysing genetic mutations, and in particular single nucleotide polymorphisms (SNPs) and/or somatic mutations, is described, as well as methods for preferentially amplifying one allelic form compared with another form. The methods use an oligonucleotide probe which hybridises to a first allele with a lower melting temperature (Tm) than that with which it hybridises to a second allele, together with amplification primers which flank the oligonucleotide probe binding site and which bind to the sample with a higher Tm than that of the probe and the first allele. An amplification reaction may be carried out at a temperature such that the probe is preferentially hybridised to the second allele, thereby amplifying the first allele. The amplified sequences may be detected using the same probe as acted as the blocking probe during amplification.

Abstract: The invention relates to devices and methods for nanopore sequencing. The invention includes compositions and methods of nucleic acid sequencing using a single polymerase enzyme complex comprising a polymerase enzyme and a template nucleic acid attached proximal to a nanopore, and nucleotide analogs in solution comprising charge blockade label that are attached to the polyphosphate portion of the nucleotide analog such that the charge blockade labels are cleaved when the nucleotide analog is incorporated into a growing nucleic acid and the charge blockade label is detected by the nanopore to determine the presence and identity of the incorporated nucleotide and thereby determine the sequence of a template nucleic acid.

Abstract: The present invention provides for a novel system and method for amplification and detection of nucleic acids within a microfluidic device wherein multiple nucleotides capable of priming PCR are present within the system and substantially sequestered within separate hydrogel posts therein.

Abstract: The present invention relates to a method of removing an intron contained in a gene from a eukaryotic gene, and linking only the exon sequences to prepare an expression vector comprising the linked sequences. Specifically, the invention relates to a method of preparing an expression vector containing linked exon sequences comprising amplifying exon sequences by PCR as one or more fragments from a giant fungal gene containing an intron, and linking the fragments together with a restriction enzyme-treated vector using the gap repair cloning method; a method of preparing an expression vector containing a full-length cDNA sequence by synthesizing and linking cDNA fragments from a fungal giant gene; a transformant having introduced therein an expression vector prepared by the method; a protein produced by the transformant; and a method of preparing a compound produced by the protein using the expression vector.

Abstract: Methods and devices are provided for pretreatment of a sample containing microbial cells. In some embodiments, the pretreatment of the sample is performed via the initial selective lysis, within a sample pretreatment vessel, of non-microbial cells (such as blood cells) and the subsequent centrifugation of the sample to remove the resulting debris and concentrate the microbial cells. An immiscible and dense cushioning liquid may be included, for collecting the microbial cells adjacent to a liquid interface formed by the cushioning liquid, upon centrifugation of the pretreatment vessel. After removal of a substantial quantity of the supernatant, resuspension of the collected microbial cells, and re-establishment of the liquid interface, at least a portion of the remaining suspension may be removed without substantially removing the cushioning liquid. One or more intermediate wash cycles may be performed to prior to extraction of the pretreated sample.

Abstract: The invention relates to a method of typing non-small cell lung cancer by determining RNA levels for a set of genes. The typing can be used for determining a metastasizing potential of the cancer cells. The invention further relates to a set of probes and a set of primers for typing non-small cell cancer cells.

Abstract: An attachment unit for attachment of a reaction container including a channel filled with a reaction solution and a liquid having a specific gravity different from that of the reaction solution and being immiscible with the reaction solution, the reaction solution moving close to opposed inner walls, a first heating unit that heats a first region of the channel and a second heating unit that heats a second region of the channel when the reaction container is attached to the attachment unit, a drive mechanism that switches arrangement of the attachment unit, the first heating unit, and the second heating unit between a first arrangement and a second arrangement in which a lowermost position of the channel is located within a first region and a second region, respectively, and a control unit that controls the drive mechanism, the first heating unit, and the second heating unit are provided.

Abstract: The disclosure provides microcrystals of Y receptor agonists; microcrystalline pellets of Y receptor agonists, and microcrystalline suspensions of Y receptor agonists. Pharmaceutical compositions containing these microcrystals, microcrystalline pellets, and microcrystalline suspensions have prolonged pharmacokinetic profiles making them useful for once daily or once weekly administration.

Abstract: The invention provides pipette tip columns and automated methods for the purification of nucleic acids such as plasmids from unclarified cell lysates containing cell debris as well as methods for making and using such columns. The columns typically include a bed of medium positioned in the pipette tip column, above a bottom frit and with an optional top frit.

Abstract: Methods and kits for generating circular nucleic acids in a cell-free system, and uses for the generated circular nucleic acids are provided. The methods comprise in vitro amplification of a nucleic acid template comprising a recombination site to produce tandem repeat nucleic acid sequence, and employ a recombination protein to generate the circular nucleic acids from the tandem repeat nucleic acid sequence.

Abstract: A method and apparatus are provided for forming high-yield tissue microarray blocks capable of producing 1,000 or more replicate slides. In one embodiment, a plurality of donor tissue samples are minced and suspended in separate molten liquid carriers to form a plurality of separate two-phase mixtures of solid minced tissue and molten wax. Each of the two-phase mixtures is preferably formed into elongated columns and transferred into a separate well of a microarray block. A method and apparatus are provided for forming the elongated molten wax columns with minced tissue evenly distributed along the length of each column. The apparatus for forming the array includes an extrusion mechanism for transferring the columns of two-phase mixtures into the deep wells of the microarray block. The two-phase mixture may alternately be cooled and allowed to solidify before being transferred into the microarray block.

Abstract: A reliable and highly sensitive method is provided for detecting methylation status of CpG-containing nucleic acids by nucleic acid amplification and melting curve analysis of amplification products. The methods and compositions employs a novel design of primers, CpG-containing methylation-independent oligonucleotide primers, wherein both unmethylated and methylated alleles of a CpG-containing nucleic acid can be detected by use of only one set of primers after the CpG-containing nucleic acid has been subjected to cytosine to thymine conversion of unmethylated Cytosine. The method is useful for detection of methylation status in for example cancer genes and other disease related genes, wherein methylation influences gene expression.