33 results about "Cleaved Cell" patented technology

The present invention provides compositions, methods, and kits for lysing cells, storing nucleic acids, amplifying nucleic, and analyzing nucleic acids. Among other things, the compositions, methods, and kits are suitable for one-step lysis and amplification of nucleic acid sequences of interest. In general, the compositions comprise TCEP and a non-ionic detergent, such as Triton X-100.

A cell lysis device for lysing cells or viruses, comprising a cell lysis tube having a sample inlet; a pump connected to the cell lysis tube for transferring a sample into the tube; a sealing unit for reversibly sealing a specific region of the tube; and a laser source for generating a laser is provide. Further, a method of lysing cells or viruses using the cell lysis device is provide. The method comprises introducing a sample containing cells or viruses and optionally magnetic beads to the cell lysis tube through the sample inlet; transferring the sample to a specific region in the cell lysis tube by means of the pump; temporarily sealing the region of the cell lysis tube where the sample is placed with the sealing unit; irradiating the sample with the laser; removing the sealing unit from the cell lysis tube; and discharging the sample from the cell lysis tube by means of the pump.

The invention discloses a method for extracting total DNAs of soil microorganisms at high purity. CTAB (Cetyltrimethyl Ammonium Bromide), lysozyme, protease and SDS (Sodium Dodecyl Sulfonate) act together to perform lysis of cells and simultaneously freeze and thaw the cells repeatedly; therefore, an excellent cell lysis effect is achieved; the humic acid is removed by utilizing PVP (Polyvinyl Pyrrolidone), CaC12 and BSA (Bull Serum Albumin); and the DNAs are precipitated by utilizing isopropanol so that the purity of the obtained DNAs is high. The method is simple and convenient; and without being purified, the DNAs obtained by the method can be used for subsequent PCR (Polymerase Chain Reaction) analysis and DGGE (Denaturing Gradient Gel Electrophoresis) analysis. The purity of the extracted DNAs meets the requirement for directly performing subsequent molecular biological research; the extracted DNA segments can be subjected to 16SrDNA amplification and DGGE atlas analysis; glue cutting recovery and basic group sequencing can be performed on specific stripes of the DGGE band; unknown bacteria can be identified through comparison with the conventional sequences in an RDP (Ribosomal Database Project) database or by establishing a new sequence probe; and therefore, the structural composition of the microbial community can be determined, and a foundation can be laid for researching the structural diversity and ecological functions of the soil microorganisms in the rhizosphere of the mulberry in future.

The invention relates to a method for extracting DNA from acidic mine water bed mud in a coal mine. Buffer solution elution, supersonic oscillation and membrane filtration for thallus collection are adopted in the method, and cell lysis is performed to extract DNA. The extracted DNA can be applied to such molecular biology technologies as PCR amplification, DGGE and TRFLP and the like for analyzing microbial communities in the acidic mine water bed mud in the coal mine without purification. The method has the advantages of being simple and rapid, and being capable of truly reflecting diversityof different microbial populations in the acidic mine water bed mud in the coal mine.

The invention provides a silicon bead test nucleic acid extraction kit as well as a preparation method and application thereof. The silicon bead test nucleic acid extraction kit includes a lysis solution A, a lysis solution B, a proteinase K solution, an adsorption solution, an adsorption bead suspension, a rinsing solution and an eluent, wherein the lysis solution A, the lysis solution B and theproteinase K solution are mixed to form a lysate. After the lysis solution A, the lysis solution B and the proteinase K solution are mixed in proportion, the use volume of the lysate can be increased,complete immersion of a tested material is facilitated, so that nucleic acids in cells can be fully released; and at the same time, the problems of low lysis efficiency and decomposition in the presence of light caused by interaction between the lysis solution A and the lysis solution B can be solved, the lysate lyses the cells sufficiently and efficiently, the lysis efficiency is high, the nucleic acids with a higher concentration is obtained, improvement of the quality of the nucleic acids is facilitated, and accuracy of subsequent operation is ensured.

The invention relates to preparation of a virus vector, and provides a method for promoting secretion of a recombinant virus vector to the outside of a cell. The technical scheme is as follows: in the process of cell culture or/and virus inoculation, a polysaccharide chemical substance containing sulfo groups is added into a culture medium to promote a virus vector to be secreted out of cells. The method specifically comprises the following steps: (1) cell resuscitation; (2) cell passage; (3) virus inoculation and culture; and (4) harvesting viruses. According to the method, a polysaccharide chemical substance containing a sulfo group is added in the virus amplification process, and the virus is promoted to be secreted from inside to outside of cells, so that in the virus harvesting process, cells do not need to be split, a culture supernatant is directly harvested to obtain the virus, and the harvesting process of a virus vector mainly located in the cells is simplified. The method provided by the invention is simple in process, harmful chemical substances such as a detergent are not introduced, the virus can be harvested without a complicated cell lysis step, and the method is economical and environment-friendly.

The invention provides a method for detecting interactions between elements within one or more DNA molecules within a cell, wherein the elements are not adjacent in the primary DNA sequence, the method comprising: a) providing a cell in which elements within one or more DNA molecules that are in close proximity are cross-linked; b) simultaneously lysing the cell and mechanically fragmenting the DNA molecules within the cell; c) proximity ligating the one or more fragmented DNA molecules; d) reversing the crosslinks in the ligated DNA molecules; e) sequencing the ligated DNA molecules; and f) analysing the sequencing data to detect interactions between elements within the one or more DNA molecules within the cell.

Provided herein are methods for the rapid preparation of amplifiable nucleic acids from biological samples, which can be applied to various applications, such as, for example, point-of-care diagnostics, service laboratory diagnostics, and molecular biology applications. These methods can be performed in 15 minutes or less, and preferably in 5 minutes or less. For most applications, no further purification of nucleic acids is needed.

The invention discloses a method for rapidly identifying shRNA effectiveness. The method comprises the following steps: (1) designing and synthesizing a plurality of candidate shRNAs as candidate functional shRNAs; (2) constructing a PLKO.1-shRNA recombinant vector based on a lentiviral vector; (3) in HEK293/HEK293T cells, co-transfecting the HEK293/HEK293T cells into the cells by using a cell transient transfection method; (4) after the transfection is finished for 6-8 hours, replacing a fresh culture medium for the cells and continuously culturing for 24-48 hours; (5) collecting the cells, cracking the cells, and extracting total protein; and (6) detecting the expression of a target protein by Western Blot, and identifying the effectiveness of the candidate shRNAs by comparing the expression difference of target proteins in a control group and a treatment group. According to the method, the time cost is saved by about 50%, the overall experimental workload is reduced by about 70%, and the labor cost is remarkably and greatly reduced.

The invention discloses a research method of an exosome PD-L1. The method comprises the steps of 1) co-culturing cells and non-natural sugar, and secreting the exosome containing the non-natural sugar from the cells; (2) marking a functional probe on the non-natural sugar, wherein the functional probe is provided with a first mark; (3) identifying protein on the surface of the exosome by using an aptamer, wherein the aptamer is provided with a second marker; and 4) enabling the first marker to interact with the second marker, and detecting the interaction effect, so that the glycosylation information of the exosome protein is detected. According to the method, the glycosylation of the exosome PD-L1 can be detected in situ, the operation is simple, invasive operations such as cell lysis are not needed, and the integrity and functions of the cells are kept.