208 results about "Typing methods" patented technology

The invention discloses a human leukocyte antigen HLA-A and HLA-B gene full-length sequencing method and an HLA gene sequencing and typing method. The HLA-A and HLA-B gene full-length sequencing method comprises the following steps of: a, performing PCR amplification on about 4kb full-length sequences of HLA-A and HLA-B genes by using a pair of primers respectively; and b, cloning the amplification products to a pGEM-Tea sy vector, sequencing the full-length sequences by using ten walking sequencing primers in positive and negative directions respectively, and totally obtaining 38 allele 4.3kb full-length sequences of the HLA-A and 30 allele 3.7kb full-length sequences of the HLA-B. The HLA-A and HLA-B sequencing and typing method comprises the following steps of: performing PCR amplification on typing target areas of the HLA-A and HLA-B by using two pairs of primers respectively; and performing two directional sequencing on products by using fourteen sequencing primers respectively, wherein the HLA-DRB1 and HLA-DQB1 sequencing and typing method comprises the following steps of: amplifying sequences of second and third exons of DRB1 and DQB1 by adopting group specificity primers respectively; performing two directional sequencing on the second and third exons of the DRB1 by adopting eight group specificity primers and three sequencing primers; and performing two directional sequencing on the second and third exons of the DQB1 by adopting four sequencing primers respectively.

A light guided keyboard system essentially comprised of a light emission circuit and a guide control circuit built in the keyboard; an EL lamp or other light emitting device being connected to the light emission circuit; and guide software such as spelling guide or typing method guide, application software control interface and driver being included in the computer software unit to guide typing of spelling, characters, applications, web pages, game software, and pre-schooling education programs in faster and easier operation of the keyboard.

The invention discloses a CRISPR (clustered regularly interspaced short palindromic repeat) based DNA detecting and typing method and an application thereof. The method comprises following steps: (1),target DNA is subjected to PCR (polymerase chain reaction) amplification by a pair of universal primers; (2), the amplified target DNA is cut by Cas9/sgRNA; (3), the cut target DNA is linked with DNAligase; (4), the linked target DNA is subjected to PCR amplification. According to the method, the target DNA can be subjected to specific detection and typing simply, rapidly and sensitively according to the specific identification cutting characteristics of the CRISPR technology on DNA, the method is one novel DNA detection method with high specificity and sensitivity, and the critical bottleneck problems about nucleic acid hybridization, specific PCR primer design and the like in current nucleic acid detection and typing fields are solved successfully. With the application of the method, L1 and E6/E7 genes of HPV16 and HPV18 in human cervical cancer cells are detected successfully.

Disclosed is a typing method for a circular touch-sensitive keyboard, comprising detecting a user's touch data on the circular touch-sensitive keyboard to initiate a pre-input state based on the touch data; detecting the user's swipe input data on the circular touch-sensitive keyboard; obtaining a character corresponding to the swipe input data; and outputting the character; wherein the circular touch-sensitive keyboard comprises an inner key zone and an outer key area, the outer key area comprises eight outer key zones, the inner key zone comprises eight sectors, each outer key zone comprises four sectors, each sector is assigned with a particular character, the circular touch-sensitive keyboard is provided with an input module for detecting the swipe input data. Also disclosed is a typing device for a circular touch-sensitive keyboard. The typing method and typing device of the present invention can increase typing speed.

The invention discloses a method for building a standard CYP450 (cytochrome p450) gene type database. The method comprises the steps as follows: comparing a specific sequence corresponding to mutation information of a CYP450 gene type with a standard human whole genome sequence to obtain a corresponding relation between the specific sequence and the standard human whole genome sequence; and converting the CYP450 gene type into a gene type taking the standard human whole genome sequence as a reference sequence according to the corresponding relation. The invention further discloses a database built by the method and a CYP450 gene typing and enzymatic activity identification method based on the database. The database is convenient for research of CYP450; the gene typing method provides unified standards for CYP450 gene typing and provides more accurate judgment criterions for diseases, drugs or the like; unknown mutation sites in CYP450 genes can be effectively detected; hundreds of samples can be detected at the same time; and the coverage range is comprehensive and wide.

The invention discloses an HLA (human leukocyte antigen) gene specific PCR (polymerase chain reaction) amplification primer, an HLA typing method and an HLA typing kit. A sequence of the PCR amplification primer comprises a primer sequences shown as SEQ ID No. (sequence identification number) 1-2, 3-4, 5-6, 7-8 and 9-10. The HLA gene typing method comprises the steps that the HLA gene specific PCR amplification primer is used for conducting PCR amplification and purification on sample DNA (deoxyribonucleic acid); a gene exon sequencing primer is used for conducting sequencing PCR amplification, purification and sequencing on an HLA gene amplification product, the sequence of the HLA gene amplification product is compared with a standard sequence; and the type of the HLA gene is determined. The HLA gene typing kit comprises the PCR amplification primer and the sequencing primer. A qualitative leap in aspects of detection flux, data quality, cost control and the like is achieved; data is more reliable and realer; and the problem that typing cannot be conducted when nucleotide of certain alleles is positioned outside an amplification area is solved.

Through amplification of a part of fragments of an NPY gene, and through an SNaPshot SNP typing method, provided is SNP markers related to the grass carp growth speed, wherein S1 is located at a 639th locus from an initiation codon (ATG, with an ATG first nucleotide as the first) in an intron 1 sequence of the NPY gene, and a base at the position is T or A; S2 is located at an 892nd locus from the initiation codon in the intron 1 sequence of the NPY gene, and a base at the position is C or A. The two SNP loci can be used for identification or breeding of grass carp with faster growth speed, and lay the foundation for screening and establishment of grass carp fast-growth new varieties.

The invention discloses a primer, a probe and a kit for fluorescence quantitative PCR (Polymerase Chain Reaction) detecting and typing of babesia. According to the invention, the primer sequence is as shown in SEQID (SEQuence IDentifier) NO.1, 2 and 3 and the probe is as shown in SEQIDNO.4 and 5. A typing method of the babesia comprises the following steps of: carrying out PCR amplification on the to-be-detected bacterium by using the primer and the probe to obtain Erich bacterium positive samples of 290bp (base pair) bands, wherein the fluorescence of the samples is enhanced at the wavelength of 640nm in fluorescence detection; and typing the babesia according to the melting temperature. The primer, the probe and the kit disclosed by the invention have obvious technical advantages, have very high specificity and sensitivity and can be used for conveniently detecting the babesia of important species.

The invention relates to a detection method of nucleic acid, particularly discloses a whole base sequence typing of mitochondria and a determination method thereof, specifically 26 pairs of PCR primer for mitochondria sequencing and a typing method based on the primers. The 26 pairs of PCR primer which are adopted by the invention cover the total length of mitochondria genome, wherein, primer 15-1, 15-2, 15-3, 24-1 and 24-2 are designed renewedly based on Chinese population and has the pertinence of the typing of Chinese population. The amplified fragment which corresponds to the primer 24-1 is the minimum and equals to 420bp. The amplified fragment which corresponds to the primer 22 is the maximum and equals to 1162bp. The dimensions of all PCR fragments are moderate and favorable for PCR amplification. The 26 pairs of PCR primer of the invention is utilized in the laboratory of applicant for investigating the community information of Chinese nation, and the whole sequence information of Chinese population is submitted to GENEBANK databases. The result shows that the invention can be definitely applied in the field of individual recognitions, paternity testing and gene diagnosis of the field of forensic medicine, anthropology, genetics and disease.

The invention provides a monomolecular gene typing method based on DNA paper folding probe specificity marking and application thereof. The method comprises the following steps: 1) preparing a DNA paper folding probe; 2) carrying out specificity marking on a target DNA through the DNA paper folding probe; 3) carrying out imaging observation on the marked target DNA under an atomic force microscope. Moreover, the monomolecular gene typing method can also be applied to haplotype typing of patient samples and haplotype typing of practical samples with unknown haplotype information. All in all, according to the method provided by the invention, single nucleotide polymorphism can be effectively detected, and the position of a single basic group is observed at high resolution under the atomic force microscope, so that the problem that fluorescent molecular marking is limited to optical diffraction limit under an electron microscope is successfully avoided, and the method can be widely applied to the research fields such as in situ test of chromogene variation and clinic gene diagnosis.

The invention belongs to a Sanmate resistance genotype molecular detection and SNP typing method of Fusarium graminearum, which can be used for drug resistance monitoring and drug resistance genotype diagnosis to Sanmate and other benzimidazole type bactericides of the Fusarium graminearum which can cause wheat scab. The detection method comprises the following three major steps: (1) extracting DNA of a nuclear genome of a strain to be detected; (2) applying three groups of primer pairs to carry out PCR reaction; and (3) identifying the genotype of the strain with the resistance to the Sanmate according to a PCR product electrophoretogram. By adopting the Tetra-primer ARMS PCR (Tetra-primer amplification refractorymutation system-PCR) technology, the strain with the drug resistance of theFusarium graminearum and the genotype thereof can be fast and accurately detected, and the level of the drug resistance can be further judged. The detection accuracy is above 95%.

The invention discloses a microsatellite loci marker combination which comprises 17 microsatellite loci markers, and simultaneously provides an efficient detection method based on the 17 microsatellite loci markers; the markers have high polymorphism, are not linked with each other and have proper fragments interval, and are easy to be detected simultaneously. The invention is improved in a detection technology, optimizes an experimental system by utilizing a multi-marker amplification technology and a fluorescent semi-automatic microsatellite typing method, thus establishing high flux, accurate and effective 17 cattle microsatellite marker compound amplification and a gene typing method.

The invention provides a salmonella CRISPR (clustered regularlay interspaced short palindromic repeats) sequencing typing method; according to the method, by DNA sequencing of a latest interval sequence between Salmonella CRISPR1 site and CRISPR2 site, CRISPR type of a strain to be detected can be determined, and the Serotype can be predicted. The method is simple, rapid, low-cost and high-resolution, is high in consistency in salmonella serotyping results, and low in requirements on laboratory equipment and software, a new CRISPR type comprises bacterial evolution and regional and other various information, and by combination of a fluorescence probe and other methods, real time and rapid detection can be realized, and the method has popularization and application value.

The invention provides a genome cytosine site epigenome typing method which includes the steps: 1) performing bisulfite whole genome methylation sequencing on male and female parents of samples to be tested and offspring samples to obtain male and female parent and offspring sample genome sequences; 2) comparing the male and female parent and offspring sample genome sequences with a reference genome to obtain comparison results to determine a cytosine site to be tested; 3) performing chromosome coordinate sequencing and reads replication removal on the comparison results, performing Call SNPs on 5-10bp upstream and downstream sequences of a known cytosine site through GATK2-V3.2 and accordingly distinguishing offspring allelic gene sequences; 4) comparing the obtained distinguished offspring allelic gene sequences with the male and female parent genome sequences, and finishing cytosine epigenome typing. The method finishes genome cytosine site epigenome typing for the first time and is mature in technology, low in cost and easy to operate, popularize and apply.

One general aspect provides a method for detecting pyrophosphoric acid in a sample solution with high sensitivity and high accuracy by a small sensor element, and an SNP typing method. In the general aspect, a sample solution having a volume of more than that of a measurement cavity is supplied to the measurement cavity through a flow path, so as to expose a droplet from the opening. The droplet has a shape of sphere. The shape of the sphere is maintained by surface tension generated on a surface of the droplet. At least part of the sample solution contained in the droplet is evaporated so as to increase a concentration of pyrophosphoric acid in the sample solution included in the measurement cavity.

The present invention discloses a method for detecting the presence of an enterovirus in a clinical sample. The invention additionally discloses a method for typing an enterovirus in a clinical sample. Both methods employ a set of primer oligonucleotides for reverse transcription and amplification that hybridize to conserved regions of the enterovirus genome, and that provide amplicons that include significant portions of the VP1 region that are characteristic of the various serotypes. In the typing method, the invention further provides a database consisting of nucleotide sequences from prototypical enteroviral serotypes, which is used to type the clinical sample by comparing the sequence of its amplicon with each prototypical sequence in the database. The invention additionally provides mixtures of primer oligonucleotides, and a kit for use in conducting the typing method that includes a mixture of the primer oligonucleotides,

The invention discloses an HLA (human leucocyte antigen) typing method based on a PacBio RS II sequencing platform. A collected sample is subjected to DNA extraction and then to PCR (polymerase chain reaction) amplification, PCR products are mixed to establish a 10k library, and PacBio RS II sequencing is performed; then, original data obtained by sequencing are corrected, and HLA typing is performed with software programs. Compared with existing HLA typing methods, the HLA typing method has super-high resolution and is of important value to applications such as clinical graft tissue matching, population genetics, anthropology and evolutiology, as well as basic research work.

The invention discloses a constructing method of a maternal plasma free DNA library and a typing method of parental alleles. Firstly, a group of specific primers for amplifying SNP in maternal plasma free DNA is designed, and comprises 720 pairs of primers, and the sequences of upstream and downstream primers are shown as SEQ ID NO:1-1440. Then, peripheral blood of pregnant women of 9-20 weeks is acquired for the extraction of the plasma free DNA, and a multiple composite PCR amplification technology is applied to the construction of the free DNA library; after the library is purified and quantified, computer sequencing is performed on an Ion TorrentTM platform. The same method is used for performing library preparation and sequencing on corresponding maternal cells and fetal tissue genomic DNA. According to sequencing results, the maternal cells are selected as homozygous SNP sites, the percentage concentrations of non-parental alleles of the free DNA in maternal plasma at the corresponding sites are analyzed, and the purpose of determining a paternal allele in the free DNA is achieved.

The invention relates to the technical field of molecular biology, and especially relates to a multiplex amplification system, and a next generation sequencing typing kit and typing method for 21 micro-haplotype sites. The 21 micro-haplotype sites in the multiplex amplification system are derived from 21 autosomes, are balanced in allele frequency, are mutually independent between the sites, and have the advantages of polymorphism and low mutation rate. The kit contains 53 PCR-amplified single-end specific primers shown in SEQ ID NO. 1-SEQ ID NO.53, and can detect the 21 microhaplotype sites in a same reaction system. The multiplex amplification system, the kit and the typing method provided by the invention can solve the technical problem that a current STR typing technology cannot obtainan ideal typing result, and allele loss may occur due to excessive number of SNP sites.

The invention discloses a new kind of ten STR gene loci for Y chromosome and the typing method, which is characterized in that polyacrylamide gel is utilized for electrophoresis typing; the ten STR gene loci for Y chromosome, which can be applied in forensic medicine field, are screened out with silver staining coloration method; furthermore, the allelic ladder of each locus is prepared; a PCR primer and the expansion condition of the Y chromosome STR locus are optimized; wherein the optimization to the PCR primer and the expansion condition and the preparation of allelic ladder can be standardized and simplified, which is suitable for popularization of base unit. The ten STR gene loci for Y chromosome can be applied for individual identification, paternity testing and gene diagnosis in forensic medicine, anthropology, genetics, disease and other field. The new kind of ten STR gene loci for Y chromosome and the typing method has the advantages of wide application prospect, which is particularly suitable for paternity testing in the condition of patrilateral loss such as death or missing in forensic medicine practice and individual identification of mixed stain in rape and gang-rape cases, in particular to the identification to suspect having azoospermia or oligospermia disease.