416 results about "Cell lysates" patented technology

A method for isolating nucleic acid which comprises:(a) applying a sample comprising cells containing nucleic acid to a filter, whereby the cells are retained as a retentate and contaminants are removed;(b) lysing the retentate from step (a) while the retentate is retained by the filter to form a cell lysate containing the nucleic acid;(c) filtering the cell lysate with the filter to retain the nucleic acid and remove remaining cell lysate;(d) optionally washing the nucleic acid retained by the filter; and(e) eluting the nucleic acid, wherein the filter composition and dimensions are selected so that the filter is capable of retaining the cells and the nucleic acid.Additionally, there is provided a substrate for lysing cells and purifying nucleic acid having a matrix and a coating and an integrity maintainer for maintaining the purified nucleic acid. Also provided is a method of purifying nucleic acid by applying a nucleic acid sample to a substrate having an anionic detergent affixed to a matrix, the substrate physically capturing the nucleic acid, bonding the nucleic acid to a substrate and generating a signal when the nucleic acid bonds to the substrate indicating the presence of the nucleic acid. A kit for purifying nucleic acid containing a coated matrix and an integrity maintenance provider for preserving the matrix and purifying nucleic acid is also provided. Further, there is provided a method for quantifying DNA, such as double-stranded or genomic DNA, isolated from cells, such as leukocytes to determine the numbers of leukocytes in a sample of leukoreduced blood.

The present invention provides a simple and rapid method for preparing purified transposase complexes that are highly suited for fragmenting DNA. The method includes forming transposase complexes with oligonucleotide adapters in cell lysate, then purifying the complexes from the other substance in the cell lysate. Purification is accomplished using a specific binding pair, in which one member of the pair is bound to an oligonucleotide adapter of the complex and the other member of the pair is bound to a solid substrate. The bound complexes can be immediately used in DNA fragmentation reactions to produce solid substrate-bound DNA fragments, which can be used for any number of purposes, including as templates for amplification and sequencing.

Coupling reactions, suitable for use in organic or aqueous media, are performed by contacting a 1,2,4,5-tetrazine with a dienophile. The dienophile may be covalently bonded to a protein, and the coupling reaction may be performed in biological media such as those containing cells or cell lysates. The reactions may be performed in the presence of primary amines, thiols, acetylenes, azides, phosphines, and products of Staudinger and / or Sharpless-Huisgen reactions Novel 3-substituted cyclopropene compounds and trans-cyclooctenes are exemplary dienophiles for these reactions.

A method of preparing a library of tagged nucleic acid fragments including contacting a population of cells directly with a lysis reagent having one or more protease to generate a cell lysate; inactivating the protease to generate an inactivated cell lysate, and applying a transposase and a transposon end composition containing a transferred strand to the inactivated cell lysate under conditions wherein the target nucleic acid and the transposon end composition undergo a transposition reaction.

Methods are disclosed for using paramagnetic particles to concentrate or harvest cells. Methods are also disclosed for clearing a solution of disrupted biological material, such as a lysate of cells or a homogenate of mammalian tissue. Methods are also disclosed for using paramagnetic particles to isolate target nucleic acids, such as RNA or DNA, from a solution cleared of disrupted biological material using the same type or a different type of paramagnetic particle. Kits are also disclosed for use with the various methods of the present invention. Nucleic acids isolated according to the present methods and using the present kits are suitable for immediate use in downstream processing, without further purification.

The invention provides reagents, kits and methods for detecting and / or quantifying proteins in complex mixtures, such as a cell lysate. The methods can be used in high throughput assays to profile cellular proteomes. In one aspect, the invention provides a peptide internal standard labeled with a stable isotope and corresponding in amino acid sequence to the amino acid sequence of a subsequence of a target polypeptide. In another aspect, the peptide internal standard is labeled at a modified amino acid residue and is used to determine the presence of, and / or quantitate the amount of a particular modified form of a protein.

The invention is directed to methods for the treatment of wounds. Such methods utilize novel compositions, including but not limited to amnion-derived multipotent cells (herein referred to as AMP cells), conditioned media derived therefrom (herein referred to as amnion-derived cellular cytokine suspension or ACCS), cell lysates derived therefrom, cell products derived therefrom, each alone or in combination.

The present invention provides sample collection, shipping, and storage devices and methods of using the same. These devices and methods are useful, for example, for collecting, shipping, and storing biological samples, such as blood, serum, buccal samples, tissue homogenates, or cell lysates in a dry state. The devices and methods facilitate the rapid drying of biological samples collected on the devices, thereby improving the quality of the stored sample, particularly the protein and small molecule components of the stored sample. The present invention further provides methods of recovering biological samples from such devices.

A method for extracting proteins from heterogeneous fluids by precipitation using microfluidics. The method uses an automated protocol for precipitation of proteins onto surfaces, rinsing the precipitates to remove impurities, and resolubilization in buffer for further analysis. The method is compatible with proteins representing a range of different physicochemical properties, as well as with complex mixtures such as fetal bovine serum and cell lysate. In all cases, the quantitative performance (measured using a fluorescent assay for % recovery) was comparable to that of conventional techniques, which are manual and require more time.

The invention provides for the detection and quantification of PSMA, PSMA', and other prostatic markers in serum samples as well as in other types of samples for use in differentiating prostate cancer, benign prostatic hyperplasia, and negative diagnoses. The diagnostic detection of nucleic acids, such as mRNAs, which encode prostatic markers in cell lysates and other sample sources is also provided. In addition to the multiplexed detection / quantification of these protein- and nucleic acid-based markers, the invention also includes biochips, kits and integrated systems.

Described herein is a method in which genomic nucleic acid of a cell can be separated from nucleic acid having a molecular weight that is lower than the molecular weight of the genomic nucleic acid (e.g., plasmid DNA) of the cell directly from a cell growth culture. Also described herein, a method in which genomic nucleic acid can be separated from nucleic acid having a molecular weight that is lower than the molecular weight of the genomic nucleic acid in a cell lysate without the need to prepare a cleared lysate. The present invention is also directed to a method of isolating genomic nucleic acid (e.g., RNA, DNA) of a cell or an organism.

A differential extraction system is directed to a microfluidic-based integrated cartridge to automate differential extraction of specific cell types within a mixed sample. The integrated cartridge includes a sonication module for selective cell lysis, separating means to eliminate centrifugation, high surface area pillar chip modules to purify DNA from a cell lysate, and microfluidic circuitry to integrate the steps in an automated platform.

Described is a method of producing a repertoire of random peptides on the surface of AAV particles wherein said random peptides are expressed as a fusion with an AAV capsid protein of an AAV particle which displays at its surface said random polypeptides. Also described is a peptide library obtainable by said method as well as a method of selecting a gene therapy vector specific for a desired cell type comprising the steps of (a) infecting the desired cell type with a peptide library of the invention and (b) harvesting AAV library particles from the supernatant and / or cell lysates. Finally, AAV vectors obtained by said method are described which are useful for gene therapy, e.g., AAV vectors targeting primary human coronary artery endothelial cells which are suitable for the treatment of diseases associated with a dysfunction of said cells.

The invention relates to a rapid vaccinia antibody detection device, method and test kit for the detection of vaccinia antibody in a serum, plasma or whole blood test sample utilizing a purified vaccinia cell lysate as the capture antigen. The detection device operates on the basis of a 2-step flow-through format in use with a push buffer.

The present invention comprises devices and methods for performing channel-less separation of cell particles by dielectrophoresis, DC high voltage-pulsed electronic lysis of separated cells, separation of desired components from crude mixtures such as cell lysates, and / or enzymatic reaction of such lysates, all of which can be conducted on a single bioelectronic chip. A preferred embodiment of the present invention comprises a cartridge (10) including a microfabricated silicon chip (12) on a printed circuit board (14) and a flow cell (16) mounted to the chip (12) to form a flow chamber. The cartridge (10) also includes output pins (22) for electronically connecting the cartridge (10) to an electronic controller. The chip (12) % includes a plurality of circular microelectrodes (24) which are preferably coated with a protective permeation layer. Specific cells from various cell mixtures were separated, lysed, and enzymatically digested on the chip.

The entire process of reverse transcription-polymerase chain reaction (RT-PCR) is simplified by using oligonucleotide-immobilized microplates made of, e.g., polypropylene, to which oligonucleotides are securely immobilized and which can be subjected to thermal cycles of PCR. RT-PCR is preferably conducted in solid-phase. Capturing of mRNA and RT-PCR can be conducted in the same plates. The cDNA synthesized from the mRNA captured on the microplates can be used more than once. Further, in combination with the microplates, a filter plate is used for the preparation of cell lysates wherein target cells are placed on the filter plate, and a lysis buffer is passed through the cell layer on the filter to transfer cell lysate directly to the microplate via well-to-well communication.

The invention provides pipette tip extraction columns for the purification of a DNA vector from un-clarified cell lysate containing cell debris as well as methods for making and using such columns. The columns typically include a bed of extraction media positioned in the pipette tip column, above a bottom frit and with an optional top frit.

The inventive self-contained apparatus for isolating nucleic acid, cell lysates and cell suspensions from unprocessed samples apparatus, to be used with an instrument, comprises at least one input, and: (i) a macrofluidic component, comprising a chamber for receiving said unprocessed sample from a collection device and at least one filled liquid purification reagent storage reservoir; and (ii) a microfluidic component in communication with said macrofluidic component via at least one microfluidic element, said microfluidic component further comprising at least one nucleic acid purification matrix; and (iii) a drive mechanism on said instrument for driving said liquid purification reagent, through said microfluidic element and said nucleic acid purification matrix, wherein the only inputs to said apparatus are via said chamber and said drive mechanism.

The invention provides methods, and related devices and device components, for detecting, sensing and analyzing analytes in samples. In some aspects, the invention provides methods, and related devices and device components, useful in combination with a mass analyzer for the mass spectrometric analysis of analytes derived from biomolecules in biological samples including biological fluids cell extracts, and cell lysates. Methods of some aspects of the invention utilize a thin membrane-based detector as a transducer for converting the kinetic energies of analytes into a field emission signal via excitation of mechanical vibrations in an electromechanically biased membrane by generation of a thermal gradient.

The invention relates to a process for making porous cross-linked cellulose membranes and processes for coupling a chromatography ligand to cross-linked cellulose membranes. The invention provides methods for separating a first component from a second component in a solution based upon a difference in the size of the first and second components, and methods for separating target molecules from other components in a solution comprising use of membranes obtainable by the process of the invention. The method has particular utility in separating proteins from cell lysates and cultures.

A sonication apparatus is directed to a microfluidic-based system to automate differential extraction of specific cell types within a mixed sample. The microfluidic-based system includes a sonication module for selective cell lysis, separating means to eliminate centrifugation, high surface area pillar chip modules to purify DNA from a cell lysate, and microfluidic circuitry to integrate the steps in an automated platform.

The present invention features use of evanescent field sensing to detect or evaluate unpurified, unlabeled analytes in raw samples. Capture molecules capable of binding to an analyte of interest can be immobilized to a sensing surface of an evanescent field sensor. The interaction between the analyte of interest in a raw sample and the immobilized capture molecules alters the refractive index at the sensing surface, thereby producing a detectable signal in the reflected light from the surface. Raw samples amenable to the present invention include, but are not limited to, cell lysates, tissue extracts, bodily fluid, or biologically-derived samples. Evanescent field sensors suitable for the present invention include, but are not limited to, grating-coupled waveguides.

Microfluidic systems and methods are disclosed which are adapted to transport and lyse cellular components of a test sample for analysis. The disclosed microfluidic systems and methods, which employ an electric field to rupture the cell membrane, cause unusually rapid lysis, thereby minimizing continued cellular activity and resulting in greater accuracy of analysis of cell processes.

The invention discloses a kit for methylation of colorectal cancer related gene multi-sites and application of the kit. A primer combination and a probe can specifically detect the methylation degrees of a plurality of colorectal cancer related gene sites. The invention further provides application and a method of the primer combination and the probe for detecting the methylation in colorectal cancer and precancerous polyps screening. The method comprises the steps that a cell lysis solution is added to cell sediment and mixed uniformly, then PVPP is added for crude extraction, a cell sediment sample after primary treatment contains a little protein and RNA, protease K and RNA enzyme are added for completely removing the protein and the RNA, cell lysate is added to a centrifugal column with an adsorption film, after impurities are washed by a DNA rising solution, a DNA eluent is added to elute the DNA in the absorption film, and finally a DNA solution is collected through centrifugation. Efficient pipe collection detection is performed aiming at the plurality of gene sites, the flexibility and the specificity are improved, and the tumor early screening requirement can be improved.