44 results about "Immobilized Oligonucleotides" patented technology

Ligation methods for manipulating nucleic acid stands and oligonucleotides utilizing hybridization arrays of immobilized oligonucleotides. The oligonucleotide arrays may be plain or sectioned, comprehensive or non-comprehensive. The immobilized oligonucleotides may in some cases be binary oligonucleotides having constant as well as variable segments. Some embodiments include amplification of ligated products.

The present method is directed to using a mutation scanning array to identify mismatches or polymorphisms in multiple genes or the same gene in multiple individuals. The array can be a chip or a microsphere. Preferably, the array has elements containing immobilized oligonucleotides that collectively span at least 10 different whole genes.

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.

Hydrazino, oxyamino and carbonyl-based monomers and methods for incorporation into oligonucleotides during enzymatic synthesis are provided. Modified oligonucleotides are provided that incorporate the monomers provided herein. Immobilized oligonucleotides and oligonucleotide conjugates that contain covalent hydrazone or oxime linkages are provided. Methods for preparation of surface bound oligonucleotides are provided. Methods for the preparation of oligonucleotide conjugates are also provided.

Particular aspects relate to a method for determining the methylation pattern of a polynucleic acid, comprising: a) preparing a solution comprising a mixture of fragments of the polynucleic acid; b) coupling the fragments with a substance being detectable with a detection method; c) contacting a solution comprising the fragments of b) with a DNA microarray having a plurality of different immobilized oligonucleotides, each comprising at least one methylation site, at respectively assigned different locations thereon, the contacing under conditions affording hybridization of fragments with correlated immobilized oligonucleotides under defined stringency, and wherein the immobilized oligonucleotides have a length of less than 200 bases; d) optionally performing a a washing step; and e) detecting, using the physical detection method, such immobilized nucleic acids to which solution fragments are hybridized and/or to which solution fragments are not hybridized.

A substrate for use in supporting high density biological or chemical arrays that is made from borosilicate or boroaluminosilicate glass. It has been demonstrated that a functionalized coating used to immobilize oligonucleotides for example, retains its functionality when exposed to environmental stresses when it is applied to a slide composed of a glass material having a low sodium oxide content.

The invention provides a nucleic acid hybridized platform based on micro-flow control and a hybridization analysis method thereof; the platform comprises an upper layer and a lower layer; the upper layer is a centrifugal micro-flow controlled chip, and the lower layer is a substrate of fixed oligonucleotide probe; the upper and the lower layers are sealed to form a fluid passage or a chamber; the hybridization analysis method takes a centrifugal force and a capillary tube force as driving forces to accelerate liquid flow to flow into a hybridized passage automatically in reciprocating manner; the chip is rotated to produce the centrifugal force, and the sample liquid flow collapses under the effect of the centrifugal force; the liquid flows through the hybridized passage and enters a temporary liquid collecting tank; then the chip is stopped rotating and the sample liquid flow is returned back to the hybridized passage from the temporary liquid collecting tank under the effect of the capillary tube force; and the repeated steps of rotating and stopping chip can reciprocate the liquid flow. The invention has the advantages of fast material transmission, short hybridizing time and low consumption of sample; and multiple samples can be analyzed in parallel simultaneously.

The invention provides a microfluidic-based nucleic acid hybridization reaction platform and a microfluidic-based nucleic acid hybridization analysis method. The platform consists of an upper layer and a lower layer, wherein the upper layer is a microfluidic chip controlled by a micropump and a microvalve; the lower layer is a substrate for fixing an oligonucleotide probe; and a fluid channel or a chamber is formed by the upper layer and the lower layer through irreversible sealing. The hybridization analysis method comprises a step of promoting liquid flow to automatically flow to and fro in a hybridization channel by taking positive pressure and negative pressure which are controlled by a program as a driving force, namely filling a sample into a microchannel by a capillary force or negative pressure after the sample enters a waste liquid chamber, and promoting liquid to flow to and fro in the microchannel by controlling the pushing and bounce (positive pressure and negative pressure) of two valve seats at both ends of the liquid channel. The microfluidic-based nucleic acid hybridization reaction platform and the microfluidic-based nucleic acid hybridization analysis method have the advantages that: mass transfer of materials is quick, hybridization time is short, the sample consumption is low, a hybridization signal of the sample can be detected in real time, the integration is easy to realize, a plurality of samples can be subjected to parallel analysis synchronously and the like.

The invention discloses an aptamer screening method of a no-fixed point target substance in the technical field of nucleic acid aptamer screening in analytical chemistry. The method comprises the following steps of: screening in a systematic evolution of ligands by an exponential enrichment (SELEX) method by building a random oligonucleotides library, and an upstream primer, a downstream primer and a fixed oligonucleotides library primer for polymerase chain reaction (PCR) proliferation and utilizing an agarose particle fixed oligonucleotides library with a streptavidin mark to obtain a PCR product for clone sequencing; and obtaining aptamers bonded with the no-fixed point target substance with the special affinity after the sequencing is finished. The method is applied to screening of heavy metal cadmium ion aptamers, so that 13 aptamers which are bonded with the cadmium ion with the high specificity and high affinity are obtained; and therefore, the aptamer screening method is good in fixing effect, high in adaptability, low in cost and suitable for aptamer screening of the no-fixed point target substance.

A substantially water-soluble polymer comprises a homopolymer or a copolymer and a first subunit precursor. The first subunit precursor comprises a first nucleic acid and an ethylene-containing moiety. The first subunit precursor is either covalently or non-covalently linked to the homopolymer or copolymer. The ethylene group is preferably tethered to the 3′- or 5′-hydroxyl position of the first nucleic acid. The substantially water-soluble polymer may further comprise a tissue-specific targeting moiety and/or a bioactive compound encapsulated by the homopolymer or copolymer.

The invention relates to gene chip used for detecting functional central nervous damage diagnosis and the preparing method. said gene chip is equipped with oligonucleotide gene probe array, the oligonucleotide biomolecule of said probe array is the special biological gene factor playing improtant role in multiple central nervous system damage process simutaneously, relating to netural cell death and tissue necrosis, inflammation, irritated response, ion dysequilibrium inside cell, signal transmission, abnormal control of cell cycle, abnormal cell construction and nerve degeneration damage morbid physiology and molecular biology active gene, respectively. The gene chip can be used to check the conditioning way, expression level and change rule of biological factor relative to cerebral ischemia damage inside body in the beginning of clinic for high risk group of cerebral apoplexy, and to predict possibility of cerebral apoplexy; and used to conditioning way and change rule of special biological factor relative to nerve damage inside nerve cell after central nervous system damage, and relationship between cell death and disease occurancy, and provides checking criterion for cilinical treatment.

Hydrazino, oxyamino and carbonyl-based reagents and methods for incorporation into oligonucleotides during their solid phase synthesis are provided. Modified oligonucleotides are provided that incorporate the reagents provided herien. Immobilized oligonucleotides and oligonucleotide conjugates that contain covalent hydrazone or oxime linkages are provided. Methods for preparation of surface bound oligonucleotides are provided. Methods for the preparation of oligonucleotide conjugates are also provided.

The invention discloses a gene chip used for researching the action of an AD (Alzheimer Disease)-controlling medicament. An oligonucleotide gene probe array is fixed on the chip, wherein the oligonucleotide is an important biomolecule gene closely related to the occurrence and development process of the AD and relates to pathophysiological molecules, such as familial AD genes and susceptibility genes, amyloidosis, microtubule and cytoskeleton recombination, cynapse function, neurotransmission and ion channel, nerve regeneration, immunoregulation, immune response, signal transduction, chondriosome, protein metabolism, and the like. The chip of the invention can be used for researching pharmacological actions and action mechanisms of the AD-preventing and treating medicament containing a chemical medicament, a biotech drug, a single traditional Chinese medicine, a traditional Chinese medicine compound and an extract thereof, researching the sifting, the prescription, and the like of the AD-controlling medicament and researching the AD pathogenesis and carrying out auxiliary and differential diagnosis on a clinical case. The gene chip overcomes the defect of researching a multifactorial disease medicament by utilizing the method of a single target and has favorable application prospect.

The present invention provides solid supports (e.g., glass) and polymer hydrogels (particularly polymer hydrogel arrays present on a solid support) comprising one or more reactive sites for the attachment of biomolecules, as well as biomolecules comprising one or more reactive sites for attachment to solid supports and polymer hydrogels. The invention further provides novel compositions and methods for the preparation of biomolecules, solid supports, and polymer hydrogels comprising reactive sites. The invention also provides for preparation of crosslinked solid supports, polymer hydrogels, and hydrogel arrays, wherein one or more biomolecules is attached by means of the reactive sites in a photocycloaddition reaction. Advantageously, according to the invention, crosslinking of the hydrogel and attachment of biomolecules can be done in a single step.

A fluorescence analyzing method includes the steps of irradiating a board, to which oligonucleotide is fixed, with light for fluorescence measurement; focusing produced fluorescence to form an image; and detecting the fluorescence with a two-dimensional sensor. Here, the board is provided with plural regions to which the oligonucleotide is fixed, and the plural regions are spaced apart from one another on the board substantially equidistantly in the vertical and horizontal directions. A fluorescent image is detected in a condition where the following equation is satisfied:

where ds denotes the interval between the regions, M denotes the imaging magnification of an optical focusing/imaging system, dd denotes the pixel pitch of the two-dimensional sensor, and n denotes an integer (n=1, 2, 3, 4, 5).

The method is used for separating nucleic acids and other similar constructs. It involves selective introduction, enhancement, or stabilization of affinity handles such as single-strandedness in the undesired (or desired) nucleic acids as compared to the usual structure (e.g., double-strandedness) of the desired (or undesired) nucleic acids. The undesired (or desired) nucleic acids are separated from the desired (or undesired) nucleic acids due to capture by methods including but not limited to immobilized metal affinity chromatography, immobilized single-stranded DNA binding (SSB) protein, and immobilized oligonucleotides. The invention is useful to: remove contaminating genomic DNA from plasmid DNA; remove genomic DNA from plasmids, BACs, and similar constructs; selectively separate oligonucleotides and similar DNA fragments from their partner strands; purification of aptamers, (deoxy)-ribozymes and other highly structured nucleic acids; Separation of restriction fragments without using agarose gels; manufacture recombinant Taq polymerase or similar products that are sensitive to host genomic DNA contamination; and other applications

A method of analysing a polynucleotide target involves incubating the target with an oligonucleotide probe, generally an array of immobilized oligonucleotide probes, to form a duplex, and using ligase or polymerase to extend one chain of the duplex. A point mutation or variable number tandem repeat section may be analyzed. Arrays of immobilized oligonucleotides are provided for use in the method.