68results about How to "Low mismatch rate" patented technology

The invention provides a primer set, a kit and a detection reaction system for detecting PKD1 gene mutation through the long fragment PCR and high-throughput sequencing technology, a non-diagnosis-purpose method for external PKD1 gene mutation detection, a non-diagnosis-purpose method for external PKD1 gene analysis and a method for detecting new mutation sites on the PKD1 gene. According to the method, the primer set is used for carrying out long fragment PCR amplification on the PKD1 gene of a sample, and detecting or analyzing is carried out through high-throughput sequencing. The autosomal dominant genetic polycystic nephrosis (adult polycystic nephrosis) can be diagnosed through the assistance of a detection result, previous unknown new mutation on a plurality of PKD1 real genes can be obtained and supplied to doctors or researchers so that the relevance between the mutation and the adult polycystic nephrosis can be studied.

The invention discloses a method for detecting pathogens of infectious diseases, which possibly exist in a biological sample. The pathogens of the infectious diseases comprise avian influenza virus H5 subtype, avian influenza virus H7 subtype, SARS coronavirus, hanta virus, plague yersinia pestis and bacillus anthracis. The method comprises amplifying a nucleic acid fragment of a biological sample and detection by a probe. The invention also provides a primer for amplification and a probe for detection. The invention also provides a kit including the primer. The method has the advantages of high flexibility, strong specificity, easy operation, wide sample range, the detection for the pathogens of various infectious diseases at the same time and the suitability for early diagnosis of respiratory infectious diseases.

The invention discloses a mixed type hot-start DNA polymerase composition, a PCR amplification kit, and applications thereof. The composition comprises Taq DNA polymerase, an anti-Taq antibody, and mutant type KOD DNA polymerase. The anti-Taq antibody is capable of specifically being combined with Taq DNA polymerase. The 147th site of wild type KOD DNA polymerase is histidine, and the 147th site of mutant type KOD DNA polymerase is lysine. The experiment results show that the mismatch rate is low when the mixed type hot-start DNA polymerase composition is used for PCR amplification, the composition has the dual characteristics of high efficient amplification and high fidelity and is heat resistant in amplification, and the amplification effect is good no matter for a long DNA segment, a template with a low amount, or a template with a high GC content.

The invention discloses expression and preparation methods for a polyvalent multi-specific antibody and an immune hybrid protein and belongs to the technical field of biology. According to the invention, by using characteristics of protein Intein, a novel method for preparing polyvalent specific antibodies, or antibody and cell factor merged hybrid immunoproteins, or antibody and toxin protein merged immunotoxins or antibody and other active protein merged immune hybrid proteins is designed and developed. According to the methods, products, i.e., the polyvalent specific antibody and the immune hybrid protein are prepared through expressing all parts of the hybrid protein separately in appropriate procaryotic or eucaryotic cell systems, carrying out high-performance affinity-chromatography purification and separation, and then, carrying out splicing in vitro under the condition of intein splicing. The methods have high production efficiency, are wide in applicable application range and facilitate the separation and purification of the products. The invention provides a novel method for preparing biological drugs for directional attack on cancers or other diseases.

The invention provides DNA single-molecule sequencing system and apparatus based on multicolor-fluorescence reversible termination nucleotide. The DNA single-molecule sequencing system comprises a primer, a DNA template to be tested and multicolor-fluorescence reversible termination nucleotide reagents. The primer is fixed on a surface of a flow cell reactor; after hybridization of the sequencingprimer with the DNA template to be tested, the primer is extended by using the multicolor-fluorescence reversible termination nucleotide reagents; and thus, sequence information of the DNA template tobe tested can be obtained by detecting fluorescence signals of the extended primer. Localizing fluorescent marker is not required for the 3'-terminal of the DNA template to be tested. By extending fluorescence of a reactant as localizing fluorescence of the next extension in sequencing cycle or adopting a localizing fluorescent marker fixed on the surface of the flow cell reactor as localizing fluorescence, the DNA single-molecule sequencing system requires no localizing fluorescent marker at the 3'-terminal of the DNA template to be tested; so that, the problem of location information loss caused by quenching is effectively avoided. Thus, sequencing reading length can be further and greatly extended with error rate reduced.

The invention discloses expression and preparation methods for a bivalent bispecific antibody and a hybrid protein and belongs to the technical field of biology. According to the methods, the aim of preparing products, i.e., the bivalent bispecific antibody and the immune hybrid protein thereof are prepared through expressing all parts of the bivalent bispecific antibody and the immune hybrid protein thereof separately in appropriate procaryotic or eucaryotic cell systems, carrying out high-performance affinity-chromatography purification and separation, and then, carrying out splicing in vitro under the condition of intein splicing. The methods have high production efficiency, are wide in applicable application range and facilitate the separation and purification of the products. The products prepared by using the methods have biological activities, i.e., bivalent bispecific immunoaffinity and cytotoxicity. The methods are novel methods for preparing biological drugs for directional attack on cancers or other diseases.

The invention discloses a method for detecting infectious disease pathogens possibly existing in a biological sample. The infectious disease pathogens comprise EB virus, herpes simplex virus, cytomegalic inclusion disease virus and adenovirus. The method comprises the following steps: expanding nucleic acid fragments of the biological sample, and detecting the nucleic acid fragments by using a probe. The invention also provides primers used for expanding and the probe used for detection. The invention also provides a kit comprising the primers. The method has the advantages of high sensitivity, strong specificity, simple operation and wide sample range, can simultaneously detect various infectious disease pathogens, and is suitable for early diagnosis of respiratory infectious diseases.

The invention device a real-time detecting device of material density on the jigger layer, which consists of ª†-ray detector along the height distribution and back-end disposing circuit. The device provides a reliable detector of jigger inner layer density distribution condition, which realizes the visible statue of bed density distribution automatically.

The invention relates to a mutation relaxation buffer layer for an InGaAs probe with a high In component. An InAs mutation relaxation layer extends on a semiconductor substrate; an arsenide variation structural material of which the In component extends on the InAs mutation relaxation layer and gradually changes in a reversed direction is used as a buffer layer. According to a research method of the InGaAs infrared probe which is arranged on an expandable semiconductor substrate and is provided with the wavelength of more than 1.7 microns, the arsenide variation buffer layer structure of which the In component gradually changes in the reversed direction is adopted, the strain in a larger mismatch InGaAs material can be better released in an expected manner, so that the defect density in an InGaAs absorption layer is reduced, and the device performance is improved; the greater freedom degree is led into a structural designing method of the InGaAs infrared probe with the wavelength of more than 1.7 microns, and the mutation relaxation buffer layer has wide application prospect.

The invention discloses a method for detecting infectious disease pathogens possibly existing in a biological sample. The infectious disease pathogens comprise chlamydia psittaci, pneumocystis carinii, leptospira, Q fever rickettsiosis and brucellosis. The method comprises the following steps: expanding nucleic acid fragments of the biological sample, and detecting the nucleic acid fragments by using a probe. The invention also provides primers used for expanding and the probe used for detection. The invention also provides a kit comprising the primers. The method has the advantages of high sensitivity, strong specificity, simple operation and wide sample range, can simultaneously detect various infectious disease pathogens, and is suitable for early diagnosis of respiratory infectious diseases.

The invention provides an NASBA method for detecting a tomato spotted wilt virus (TSWV). The sequences of NASBA primers for detecting the TSWV are SEQ ID No: 1-2 respectively. According to the highly conserved region of gene N of the TSWV, two inner primers with specificity are designed. The conserved gene sequences are shared by different strains with TSWV to ensure the reliability in detecting different sources of TSWV at the level of strains. The NASBA method is suitable for rapid detection and confirmation of TSWV and can be widely used in disease monitoring in production and environment, as well as TSWV confirmation in the import and export trade.

The invention discloses a library building method for sRNA sequencing. The library building method comprises the following steps that firstly, an obtained sample is pretreated, and RNA is extracted and prepared; then, a 3'end connector and a 5 'end connector which are subjected to pre-adenylation are pretreated, and the 3' end connector and the 5 'end connector are connected with the prepared RNA and purified; reverse transcription reaction is carried out on the purified connection product, meanwhile, the obtained corresponding cDNA chain is purified, and PCR enrichment reaction is carried out on the cDNA chain to obtain a PCR product containing adapter; and finally, the PCR products are subjected to size sorting and purification according to the requirements of the sRNA library, the sRNA high-throughput sequencing library is obtained, and the mismatch rate is reduced.

The invention discloses a method for constructing a genetic engineering cell strain of an obesity-resistant medicine target point UCP1, and besides, discloses establishing and application of an obesity-resistant medicine high-flux screening model. Mainly a CRISPR / Cas9 system is related and utilized, two unique sgRNAs are designed, luciferase-T2A-tdTomato-WPRE-pA is knocked in after N end ATG of aUCP1 gene of cells, particularly luciferase and tdTomato are knocked in gene sites of immortalization brown fat cells UCP1, a first stably-transfected brown fat cell strain in which luciferase and tdTomato are inserted in a promoter region of the UCP1 is formed, and the first stably-transfected brown fat cell strain is applied to high-flux screening of obesity-resistant medicines, evaluation of the obesity-resistant medicines, evaluation of obesity-resistant active substances of organisms and development of a UCP1 detection reagent kit. The UCP1 is uncoupling protein specifically expressed atbrown fat tissue, and is an obesity-resistant new target point. The construction of the stably-transfected genetic engineering cell strain has important significance in the aspects of applying reportgenes and a high-connotation method, performing high-flux screening on a compound library acutely, accurately and efficiently, and obtaining obesity-resistant medicines capable of promoting thermogenesis and reducing weight.

The invention relates to the technical field of solid-liquid-gas three-phase separating and sorting equipment and in particular to a power-function-shaped conical swirler. The power-function-shaped conical swirler is composed of a feeding pipe, a cylindrical barrel, an overflow pipe, a conical pipe and a bottom flow pipe. The feeding pipe is connected with the outer wall of the cylindrical barrelin a tangent line or involute or spiral line or other line shape feeding mode, the overflow pipe is arranged in the middle of the upper end of the cylindrical barrel, the lower end of the overflow pipe stretches into the cylindrical barrel, and the conical pipe is arranged at the lower end of the cylindrical barrel. The power-function-shaped conical swirler is characterized in that the conical pipe is arranged to be a round pipe with the perpendicular section in a power function shape, the upper end of the round pipe in the power function shape is connected with the lower end of the cylindrical barrel, and a bottom flow opening is formed in the lower end of the round pipe in the power function shape and is connected with the bottom flow pipe. The power-function-shaped conical swirler has the advantages that the sorting space of the lower portion of the swirler cone is enlarged, a zero-speed enveloping surface section is optimized, the mismatching probability of coarse and fine particles of materials in the swirler is reduced, and the overflow particle size composition of the swirler is improved.

The invention provides application of type 6 collagen in preparation of related drugs and nerve grafts capable of improving regeneration nerve orderliness. The type 6 collagen comprises human-derivedtype 6 collagen or animal-derived homologous type 6 collagen, and the animal-derived homologous type 6 collagen comprises type 6 collagen of rat, mouse or bovine. According to the invention, the type6 collagen COL6 is used as hydrogel to be filled in a nerve conduit, so that the clustering and orderliness of regenerated nerve fibers can be promoted, the regenerated nerve fibers are more close toa normal nerve tract structure in morphology, the mismatch probability of nerves and target organs is reduced, the nerve repair effect is improved, and the protein source is wide.

The invention provides a high-fidelity Pfu DNA polymerase mutant. The high-fidelity Pfu DNA polymerase mutant is characterized in that on the basis of a wild type Pfu DNA polymerase, mutation of one, three or six amino acid sites is carried out, and the mutation sites are selected from the following sites: Y403A, P411H, V438K, A741S, R757Q and S768K. The invention also discloses coding DNA of the high-fidelity Pfu DNA polymerase mutant and application of the high-fidelity Pfu DNA polymerase mutant in NGS. The high-fidelity Pfu DNA polymerase mutant has ultrahigh fidelity and high amplification efficiency, can be stably and efficiently applied to the library enrichment process in next-generation sequencing, effectively improves the library sequencing quality, reduces the base mismatch rate, and can be used for precise analysis and interpretation of genetic information.

The invention discloses a buffer solution and application thereof. The buffer solution is prepared from a Tris buffer solution with a concentration of 33 to 66 mM, divalent cations with a concentration of 1.6 to 5 mM, monovalent cations with a concentration of 25 to 75 mM, dithiothreitol (DTT) with a concentration of 0.5 to 10 mM, PEG 4000 to 8000 with a concentration of 5 to 15% and ATP with a concentration of 0.5 to 2 mM. The pH value of the Tris buffer solution is 7 to 9. The buffer solution can improve the efficiency of a ligation reaction, increase the conversion rate of a substrate and reduce the mismatch rate of the ligation reaction, is good in compatibility and small in inhibition effect on ligase, and remarkably reduces the enzyme consumption of a system, so experiment cost is reduced.

The invention discloses a follicle-stimulating hormone receptor activity detection kit and a detection method thereof. The detection kit comprises two pairs of amplification primers and a multiplex PCR mixed reaction solution, wherein the amplification primers are used for amplifying obese gene polymorphic sites and comprise rs6165 sites and rs6166 sites, and each pair of primers respectively corresponds to upstream and downstream regions of one SNP site. According to the follicle-stimulating hormone receptor activity detection kit and the detection method thereof, a multiplex PCR amplification method is adopted, two important SNP sites having high relevancy with FSHR activity are amplified at the same time, the amplified site combination can be flexibly selected according to actual needsto achieve the function of one kit with multiple functions, and primers in the kit are high in specificity and low in mismatch rate. The primers are reasonable in length and Tm value design, and the amplified product is convenient to detect and isolate. Multiple target products are amplified once, so that the amplification efficiency can be greatly improved, and the amplification cost can be reduced. The detection kit can be extended to other SNP site detection, and has a good popularization value.

The invention discloses a medicine dispensing and distributing system. The system comprises a hospital prescription subsystem used for receiving prescription information, generating a corresponding prescription instruction and transmitting the prescription instruction to a medicine distribution subsystem, wherein the prescription instruction comprises the name of a patient corresponding to a prescription, the taking number of the prescription, the name of each traditional Chinese medicine formula granule required by each prescription, the corresponding number and the storage position thereof;a medicine distribution subsystem used for receiving a prescription instruction from the hospital prescription subsystem, executing a medicine distribution task according to a pre-configured program,and sending a task completion signal after medicine distribution is completed. According to the medicine dispensing and distributing system, automatic medicine dispensing is achieved through cooperative work of all the system modules, and compared with a previous manual medicine dispensing mode, the medicine dispensing efficiency is greatly improved, the mismatching rate is reduced, and the medical service level of a hospital can be improved.

The invention discloses a method for detecting and identifying Lactobacillus acidophilus NCFM. Fragments of to-be-detected strain 16SrDNA are subjected to PCR amplification by means of primers plb16A,mlb16A, plb16B, mlb16B, plb16X, mlb16X and high-fidelity DNA polymerase; an amplification product is digested by restriction endonucleases AhdI, BmrI and XcmI, purified and connected with a T vector;a connecting product is transformed into Escherichia coli, connexons are screened, plasmids are extracted for sequencing and comparison, and whether the detected strain is the Lactobacillus acidophilus NCFM is identified preliminarily. The method has the following advantages: the Lactobacillus acidophilus can be identified by means of the length of the amplification product; the Lactobacillus acidophilus can be identified by means of the length of the amplification product after being digested by AhdI, BmrI and XcmI; the amplification product is obtained through amplification by high-fidelityDNA polymerase, so that the mispairing rate of the amplification product is low, sequencing and comparison are performed after the amplification product is connected with the T vector, and the Lactobacillus acidophilus NCFM can be identified preliminarily by means of a result.

The invention discloses a quick amplifying reagent kit and amplifying method of a CYP2C19 gene. The reagent kit comprises 3 pairs of amplifying primers, Taq enzymes, dNTPs and a buffered solution, wherein the amplifying primers are used for amplifying at least two SNP sites in a CYP2C19*2 type, a CYP2C19*3 type and a CYP2C19*17 type, and each pair of primers corresponds to upstream and upstream regions of one of SNP sites. A multiplex-PCR amplifying method is used, a plurality of important SNP sites of the CYP2C19 are amplified at the same time, the combination of sites to be amplified can beflexibly selected according to actual demands, and the effect that one reagent kit has many functions can be achieved; the primers in the reagent kit are high in specificity and low in mispairing rate, the length and the Tm value of the primers are reasonable to design, and amplified products are convenient to detect and separate. Multiple purpose products are amplified at a time, so that the amplifying efficiency can be greatly improved, and the amplifying cost can be reduced.

The invention discloses a primer and a method for detecting gene mutation related to congenital deficiency of blood coagulation factor XII. The method comprises the following steps: performing amplification testing on 14 pairs of primers in a whole-exome sequence in a hot spot region of blood coagulation factor XII (F12); and adopting Sanger sequencing techniques and sequencing primers. Accordingto the primer and method disclosed by the invention, mutation of genes related to the congenital deficiency of blood coagulation factor XII can be rapidly detected. The method disclosed in the invention has accurate results, is capable of aiding diagnosis of the congenital deficiency of blood coagulation factor XII and has important reference significance on early intervention, early treatment andantenatal diagnosis.

The invention provides an MSGV recombinant carrier as well as a preparation method and application thereof. The recombinant carrier comprises a gene segment comprising a TRGC gene, a TRDC gene and a CD3-zeta gene. A C area of alpha-beta TCR on the MSGV carrier is replaced by an extracellular C area of gamma-zeta TCR as well as a transmembrane area and an intracellular area of the CD3-zeta, so thatthe mismatch rate of exogenous TCR molecules and endogenous TCR is obviously reduced.

The invention discloses an NASBA primer, a kit and a method for detecting peach latent mosaic viroid. The sequences of an upstream primer and a downstream primer are respectively SEQ ID No. 1-2. An NASBA amplification kit comprises the following components: (1) NASBA amplification reaction liquid A; (2) NASBA amplification reaction liquid B. The detection method is as follows: 1) extracting sample RNA; 2) carrying out PLMVd NASBA amplification; and 3) carrying out electrophoresis detection. The NASBA primer, kit and method disclosed by the invention are suitable for quick detection and verification of peach latent mosaic viroid and can be widely used for monitoring epidemic situations in agricultural production and environment and monitoring and detecting the kind of viruses in import and export trades, the operation is very simple and convenient, the necessary sample amount is small and the requirements on the quality of template RNA are quite low.

The invention discloses a sperm recognition and multi-target trajectory tracking method, which comprises the following steps of: selecting a target sperm from a first frame of sperm image, acquiring coordinates of the target sperm, and taking the coordinates as starting coordinate points of a trajectory chain of the target sperm; calculating a prediction coordinate point of the target sperm in theith frame of sperm image by using a trajectory prediction algorithm; setting a search area according to the predicted coordinate points in the ith frame of sperm image for searching, if a plurality of sperms are searched in the area, performing association matching by combining multiple principles with a matching algorithm to obtain an optimal matching sperm, and adding the coordinate points of the optimal matching sperm into the trajectory chain of the target sperm; and if no sperm is searched in the area, adding the predicted coordinate point into the trajectory chain of the target sperm. According to the invention, the tracking efficiency of sperm motion trails is improved, the method is suitable for the occasion of temporary loss of sperms, the matching precision is improved, and themismatch rate is reduced.

The invention discloses an MLPA-enhanced specific method for detecting SNP sites. An MLPA probe aiming at Kras gene 216G-CSNP sites is designed, an MLPA experiment is used, and the influence on the SNP sites during detection is observed. The key points of the method disclosed by the invention are that the mismatch rate of the probe on the SNP sites is reduced when the specificity is detected at the SNP sites, and the probe can be synthesized by using nucleotide subjected to specific chemical modification. The research proves that after the SNP sites of the MLPA probe are modified by using LNA, the base mismatch rate can be obviously reduced by virtue of LNA modification, and the false positive product peak is eliminated; however, the detection sensitivity of the LNA modified probe is reduced, and when a positive sample is detected, compared with a non-modified MLPA probe, the probe disclosed by the invention has the advantages that the product peak area is reduced, probe hybridization or continuous efficiency reduction can be possibly caused by LNA modification, and the detection sensitivity of MLPA is reduced.

The invention discloses a NASBA primer, a reagent kit, and a method for detecting apple scar skin viroid; the sequences of the upstream primer and the downstream primer are respectively SEQ ID NO: 1-2; the NASBA augmentation reagent kit comprises the constituents as follows: (1) NASBA augmentation reaction liquid A; (2) NASBA augmentation reaction liquid B; the detection method is as follows: 1) extracting sample RNA; 2) augmenting NASBA of ASSVd; 3) detecting with electrophoresis. The NASBA primer, the reagent kit, and the method for detecting apple scar skin viroid are applicable to quickly detecting and confirming apple scar skin viroid, can be widely applied for epidemic surveillance in agricultural production and environment as well as monitoring and detecting the viroid in import and export trade; the operation is very simple; the amount of the required sample is small; and the quality requirements to template RNA is lower.