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218 results about "Genomic Segment" patented technology

Genomic segment extracted from a library e.g. BAC or YAC library to be used in the genetic manipulation of the animal model.

Cas9 nuclease K918A and application thereof

The invention belongs to the technical field of biology and particularly relates to Cas9 nuclease and application. The Cas9 nuclease (Cas9-K918A) disclosed by the invention has the activity of the Cas9 nuclease, is suitable for a CRISPR / Cas9 system and is obtained by mutating 918th lysine of wild type Cas9 nuclease into alanine. The Cas9 nuclease (Cas9-K918A) is adopted for cutting DNA (Deoxyribonucleic Acid) double strands to generate a predominantly-broken terminal; a basic group complemented with the predominantly-broken terminal can be added in a connection filling-in mode, so that accurate edition of a specific position of a genomic DNA fragment can be realized.
Owner:SHANGHAI JIAO TONG UNIV

Specific and universal probes and amplification primers to rapidly detect and identify common bacterial pathogens and antibiotic resistance genes from clinical specimens for routine diagnosis in microbiology laboratories

The present invention relates to DNA-based methods for universal bacterial detection, for specific detection of the common bacterial pathogens Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Staphylococcus saprophyticus, Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis as well as for specific detection of commonly encountered and clinically relevant bacterial antibiotic resistance genes directly from clinical specimens or, alternatively, from a bacterial colony. The above bacterial species can account for as much as 80% of bacterial pathogens isolated in routine microbiology laboratories. The core of this invention consists primarily of the DNA sequences from all species-specific genomic DNA fragments selected by hybridization from genomic libraries or, alternatively, selected from data banks as well as any oligonucleotide sequences derived from these sequences which can be used as probes or amplification primers for PCR or any other nucleic acid amplification methods. This invention also includes DNA sequences from the selected clinically relevant antibiotic resistance genes. With these methods, bacteria can be detected (universal primers and/or probes) and identified (species-specific primers and/or probes) directly from the clinical specimens or from an isolated bacterial colony. Bacteria are further evaluated for their putative susceptibility to antibiotics by resistance gene detection (antibiotic resistance gene specific primers and/or probes). Diagnostic kits for the detection of the presence, for the bacterial identification of the above-mentioned bacterial species and for the detection of antibiotic resistance genes are also claimed. These kits for the rapid (one hour or less) and accurate diagnosis of bacterial infections and antibiotic resistance will gradually replace conventional methods currently used in clinical microbiology laboratories for routine diagnosis. They should provide tools to clinicians to help prescribe promptly optimal treatments when necessary. Consequently, these tests should contribute to saving human lives, rationalizing treatment, reducing the development of antibiotic resistance and avoid unnecessary hospitalizations.
Owner:GENEOHM SCI CANADA

Quantitative sequencing and library building method and quantitative sequencing and detecting method for fusion gene on basis of DNA (Deoxyribonucleic Acid) and application of quantitative sequencing and detecting method

The invention discloses a quantitative sequencing and library building method for a fusion gene on the basis of DNA (Deoxyribonucleic Acid). The quantitative sequencing and library building method comprises the following steps: firstly, constructing a genome fragmentation DNA library and purifying the library; secondly, capturing a fusion gene generation region by PCR (Polymerase Chain Reaction) amplification, purifying the captured gene and enriching a sequence containing a specific primer fragment; thirdly, capturing a target fragment containing the fusion gene by nested PCR amplification; fourthly, constructing a DNA sequencing library with high-throughput sequencing. The invention also discloses a quantitative sequencing and detecting method for the fusion gene by using the DNA sequencing library prepared by the quantitative sequencing and library building method, application of the quantitative sequencing and detecting method as well as a detection kit containing the DNA sequencing library. According to the quantitative sequencing and library building method disclosed by the invention, the downstream where a fusion breakpoint occurs is anchored by using a one-way specific primer; a target sequence is obtained by pairing specific primers with universal primers and using a PCR method; the background is further reduced label enriching and nested PCR, so that the specificity is improved, the time for building the library is shortened, and the cost for building the library is reduced; the quantitative sequencing and library building method is suitable for an FFPE (Formalin Fixed And Parafiin Embedded) sample or liquid biopsy.
Owner:CARRIER GENE TECH SUZHOU CO LTD +1

Virulence of streptococci

The invention relates to the field of diagnosis of and vaccination against Streptococcal infections and to the detection of virulence markers of Streptococci. The invention discloses a method for modulating virulence of a Streptococcus, the method comprising modifying a genomic fragment of Streptococcus wherein the genomic fragment comprises at least a functional part of a fragment identifiable by hybridization in Streptococcus suis to a nucleic acid or fragment thereof as shown in FIG. 5.
Owner:STICHTING WAGENINGEN RES

Method and device for assembling genomes by utilizing long transcriptome sequencing result

The invention relates to a method and a device for assembling genomes by utilizing a long transcriptome sequencing result. The method adopts the scheme that transcriptome sequencing reading segments and genome segments of the same species are compared; only one transcriptome sequencing reading segment from a comparison to one genome segment is removed, query blocks of reserved transcriptome sequencing reading segments are filtered under specified conditions; block connection relative to the reserved query blocks are obtained according to the specified conditions; and the genome segments are connected on the basis of the block connection, thereby completing assembling of the genome sequences. According to the method for assembling the genomes by utilizing the long transcription sequence result, long segment sequencing data including a large amount of existing and published Sanger data can be utilized, so that the genome sequences can be assembled by utilizing the long transcriptome sequencing reading sequence.
Owner:CHINESE ACAD OF FISHERY SCI

Quality control method for detecting human EGFR (Epidermal Growth Factor Receptor) gene variation based on high-throughput sequencing and kit

The invention discloses a quality control method for detecting human EGFR (Epidermal Growth Factor Receptor) gene variation based on high-throughput sequencing and a kit, and applications thereof. The quality control method comprises the following steps: extracting a plurality of genome DNAs of an EGFR gene variation-positive human tumor cell line; measuring variation positive sites as positive control sites through a Sanger sequencing method; fragmenting the genome DNAs and mixing according to a certain proportion to obtain a quality control product which can be applied high-throughput sequencing detection of human EGFR gene variation. The kit comprises the human EGFR gene variation detection quality control product.
Owner:3D BIOMEDICINE SCI & TECH CO LTD

Recombinant influenza virus vector carrying foreign genes in NA segment and preparation method and application thereof

The invention discloses a recombinant influenza virus vector carrying foreign genes in an NA segment, which can be rescued in cells by a virus rescue method and can be amplified in chick embryos in large quantities. The invention also discloses a preparation method thereof, comprising the following steps: firstly constructing plasmids used for rescuing influenza viruses and introduced with the foreign genes on the NA genome segment of the influenza virus and carrying out rescue on host cells. The foreign gene is connected with the NA genome segment by a 2A segment. The foreign gene comprises EGFP reporter genes and adopts a CMV / huPolI bidirectional influenza virus rescue system. The recombinant influenza virus vector overcomes capacity limit of the foreign genes in the influenza virus genome, enjoys stable passage in chick embryos, can be applied to vaccine and drug development and tumor treatment and can produce recombinant cell factor drugs in cheap chick embryo bioreactors.
Owner:GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI

DNA based fusion gene quantitative sequencing database foundation and detection method and applications thereof

ActiveCN107699957ASolve the difficulty of obtaining RNA of appropriate qualityAvoid background influenceMicrobiological testing/measurementLibrary creationSequence databaseLiquid state
The invention discloses a DNA based fusion gene quantitative sequencing database foundation method. The method comprises (1) carrying out genome DNA fragmentation; (2) sorting DNA fragments; (3) carrying out unidirectional specific PCR amplification to obtain an amplification product with fusion sites; (4) connecting a single chain general library linker sequence to the 3' terminal of the amplification product; and (5) carrying out PCR amplification, enriching fusion gene regional fragments, and carrying out purification and quantification to obtain sequencing database. The invention also discloses a method of using a DNA library established by the provided method to carry out fusion gene sequencing detection, an application thereof, and a fusion gene unidirectional specific primer set forthe database establishment method. By unidirectionally specifically amplifying the DNA segments, capturing fusion sequences, and obtaining a DNA library for NGS sequencing through enrichment, the ratio of captured effective database is increased, the database establishment and detection speed is accelerated, and the method is suitable for FFPE sample or liquid state biopsy.
Owner:CARRIER GENE TECH SUZHOU CO LTD +1

Cancer treatment with retroviral vectors comprising wild-type p53

Disclosed are methods and compositions for the selective manipulation of gene expression through the preparation of retroviral expression vectors for expressing antisense sequences, such as K-ras oncogene antisense sequences, or sequences encoding a desired product, such as wild type p53 sequences. Preferred retroviral vectors of the present invention incorporate the β-actin promoter in a reverse orientation with respect to retroviral transcription. Preferred antisense RNA constructs of the present invention employ the use of antisense intron DNA corresponding to distinct intron regions of the gene whose expression is targeted for down-regulation. In an exemplary embodiment, a human lung cancer cell line (NCI-H460a) with a homozygous spontaneous K-ras mutation was transfected with a recombinant plasmid that synthesizes a genomic segment of K-ras in antisense orientation. Translation of the mutated K-ras mRNA was specifically inhibited, whereas expression of H-ras and N-ras was unchanged. A three-fold growth inhibition occurred in H460a cells when expression of the mutated ras p21 protein was down-regulated by antisense RNA and cells remained viable. The growth of H460a tumors in nu/nu mice was substantially reduced by expressed K-ras antisense RNA.
Owner:BOARD OF RGT THE UNIV OF TEXAS SYST
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