11 results about "Oligonucleotide hybridization" patented technology

The invention provides a method which allows the separation of different workflow steps for barcoding of target nucleic acids and therefore providing optimal reaction conditions for each workflow step, especially for template switching reactions. Moreover this method provides the opportunity to perform the reactions such as barcoding reactions of two different nucleic acid molecules from one cell such as RNA and genomic DNA molecules in a single step. The method comprises the steps: (a) Providing a plurality of cells comprising target RNA molecules and at least one solid support comprising capture oligonucleotides for said target RNA molecules and barcode oligonucleotides; (b) Partitioning said plurality of cells and said solid supports such that each cell is included into a separate partition and each partition comprises a solid support; (c) Lysing said cell, thereby obtaining a mixture of target and non, target RNA molecules; (d) Hybridizing said target RNA molecules to the capture oligonucleotides for said target RNA molecules, thereby obtaining target RNA molecules attached to said solid support (e) Disrupting the partitions and separating the non-target RNA molecules from the target RNA molecules attached to said solid support (f) Generating double stranded nucleic acids from the target RNA molecules by nucleic acid synthesis, wherein the capture oligonucleotides serve as primer and the target RNA molecules serve as templates (g) Attaching the barcode oligonucleotides to the double stranded nucleic acids from target RNA molecules, thereby generating barcoded nucleic acids from target RNA molecules; Characterized in that in step a) said plurality of cells additionally comprise target genomic DNA molecules and said at least one solid support additionally comprise capture oligonucleotides for target genomic DNA molecules and in that the capture oligonucleotides for target RNA and target genomic DNA molecules are different.

The present invention provides a single-stranded antisense oligonucleotide or pharmaceutically acceptable salt thereof for regulating the expression and / or function of the ATXN7 gene. In the single-stranded antisense oligonucleotide, each nucleotide is bonded by a phosphate group and / or modified phosphate group. The single-stranded antisense oligonucleotide includes a gap region, a 3' wing region bonded to the 3' terminal of the gap region, and a 5' wing region bonded to the 5' terminal of the gap region. The gap region is a deoxyribose-constituted nucleic acid in which a nucleic acid modified by a sugar moiety may be included. The 3' wing region and the 5' wing region are modified nucleic acids. The sugar modification constituting the single-stranded antisense oligonucleotide is a modified nucleic acid represented by formula (A1). The single-stranded antisense oligonucleotide has a base length of 12-30 mer. The base sequence of the antisense oligonucleotide is: a base sequence having a sequence identity of 90-100% with a base sequence that is complementary to at least one target region constituted at the same base length as the antisense oligonucleotide in the base sequence shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5; a base sequence that is complementary to the base sequence obtained by deleting, substituting, inserting, or adding one or more bases in the target region; or a base sequence that, under stringent conditions, hybridizes with an oligonucleotide having the target region.

1. The present invention relates to methods and kits for isolating a target nucleic acid by in-gel affinity electrophoresis. The method comprises the steps of: a. covalently immobilizing an anchor oligonucleotide during polyacrylamide polymerization in a polyacrylamide binding gel, wherein the anchor oligonucleotide comprises (a) a binding element that covalently binds to acrylamide of the polyacrylamide binding gel and (b) a first coupling nucleotide sequence that is complementary to a nucleotide sequence of a bridge oligonucleotide to hybridize the anchor oligonucleotide to the bridge oligonucleotide, wherein the bridge oligonucleotide comprises (a) a second coupling nucleotide sequence that is complementary to the first coupling sequence of the anchor oligonucleotide and (b) a landing site with at least one complementary target nucleotide sequence, b. hybridization of the anchor oligonucleotide to the bridge oligonucleotide, c. performing a polyacrylamide gel electrophoresis (PAGE) to enrich target nucleic acid by in-gel hybridization, d. depletion of undesired species by in-gel nucleic acid depletion, e. isolating the hybridized target nucleic acid from the binding gel.

Aptamer detection techniques with chemistry attachment with proximity hybridization and extension are described in an aptamer-based assay. In an embodiment, capture oligonucleotides can be used such that the capture oligonucleotides attach to spatially-fixed analytes in a fixed tissue sample. Aptamers that hybridize to the capture oligonucleotides are tagged with an extension sequence that can then be used for subsequent capture and spatial tagging steps.

An improved nucleic acid capture method, comprising the following steps: by means of a splint oligonucleotide hybridization random probe and an oligonucleotide strand fixed on a capture chip, an RNA in a sample captured by the random probe undergoing reverse transcription into a cDNA, and linking the cDNA to the oligonucleotide strand, the reverse transcription and the linking being carried out in the same reaction system. During the process, while the RNA undergoes the reverse transcription into the cDNA, the cDNA is linked, by means of splint hybridization of fixed oligonucleotide sequences at two ends, to the oligonucleotide strand fixed on the chip. The method can increase the number of captured genes, and reduce the loss of cDNA; in addition, the method combines what was originally three steps into one step, thus further shortening the duration of the process.

The present invention relates to the detection of a target nucleic acid sequence by a PTOCE (PTO Cleavage and Extension) assay. The present invention detects a target nucleic acid sequence in which the PTO (Probing and Tagging Oligonucleotide) hybridized with the target nucleic acid sequence is cleaved to release a fragment and the fragment is hybridized with the CTO (Capturing and Templating Oligonucleotide) to form an extended duplex, followed by detecting the presence of the extended duplex. The extended duplex provides signals (generation, increase, extinguishment or decrease of signals) from labels indicating the presence of the extended duplex and has adjustable Tm value, which are well adoptable for detection of the presence of the target nucleic acid sequence.

A method includes inserting a fluid including a sample into a cartridge, lysing the sample in an extraction chamber to release nucleic acids of interest from the sample, collecting a first set of hybridized magnetic particles hybridized to a first type of nucleic acids of interest by: hybridizing the first type of nucleic acids of interest with an oligonucleotide attached to magnetic particles, holding the first set of hybridized magnetic particles on a side of the extraction chamber, suspending the first set of hybridized magnetic particles with a wash buffer, transporting the first set of hybridized magnetic particles to a detection chamber, collecting a second set of hybridized magnetic particles hybridized to a second type of nucleic acids of interest, amplifying the sets of hybridized magnetic particles, and detecting a plurality of amplification products indicative of the presence, absence, or amount of the types of nucleic acids of interest.

Products for detecting a plurality of protein analytes comprise a plurality of proximity probe pairs comprising first and second proximity probes having an antibody or antibody fragment specific for the same protein analyte and a nucleic acid domain, which probes can simultaneously bind to the analyte. Each pair is specific for a different analyte. Each nucleic acid domain comprises an ID sequence and at least a first hybridisation sequence. In each probe pair, ID sequences correspond to a particular analyte, and the probes comprise paired hybridisation sequences. For each probe pair, a splint oligonucleotide comprises hybridisation sequences complementary to each of the paired hybridisation sequences. When probes bind to their protein analyte, the respective paired hybridisation sequences can hybridise to the splint oligonucleotide. At least one pair of hybridisation sequences is shared by at least two pairs of proximity probes. A plurality of sample index oligonucleotides is also included.

The present disclosure provides methods and compositions, e.g., kits, for isolating eukaryotic nucleic acid from a stool sample. In one embodiment, the method provided herein involves incubating the stool sample with a chaotropic salt, heating the stool sample, and using a substrate coupled to a locked nucleic acid (LNA) oligonucleotide to isolate the nucleic acids hybridized to the LNA oligonucleotide.

Methods, kits, and systems for analyzing a sample comprising extracellular vesicles. In various examples, a method comprises use of a fluidic device for the retention and discrete presentation of single extracellular vesicles for imaging-based analyses. In various examples, a method comprises use of a plurality of capture agents that are each linked to a different oligonucleotide and a corresponding plurality of labeled nucleic acid probes, where each of the labeled nucleic acid probes specifically hybridizes with one or more or each of the oligonucleotides of the capture agents, and sub-sets of the capture agents are detected by imaging-based means using iterative cycles using corresponding sub-sets of the labeled nucleic acid probes. In various examples, kits and systems are provided for carrying out a method of the disclosure.

The present disclosure describes a system and methods for the generation of DNA strands and querying a DNA database rapidly and accurately. The method disclosed includes the generating at least one database single stranded DNA (ssDNA) strand, includes a plurality of functional oligonucleotides separated by introns. On one end of each intron there is a donor fluorophore and on the other end is an acceptor fluorophore. A query ssDNA strand is also generated that includes a series of complimentary oligonucleotides, which can hybridize with the functional oligonucleotides of the database ssDNA strand. Hybridization causes the intron regions of the database ssDNA strand to fold resulting in the donor fluorophore and acceptor fluorophore being placed in close proximity to one another. The close proximity enables Förster Resonance Energy Transfer (FRET) phenomena to occur, which is detected using a photodetector.