896results about How to "High amplification efficiency" patented technology

Methods and compositions for digital profiling of nucleic acid sequences present in a sample are provided.

Use of polymorphism site gene to predict ACEI medicine, its method and reagent kit are disclosed. It can be used to determine critical enzyme gene polymorphism site gene typing oligonucleotide and critical enzyme gene polymorphism site gene in cysteine metabolic pathway.

The invention relates to a pathogenic microorganism DNA detection chip and the chip comprises a carrier and a nucleic acid probe on the carrier. The nucleic acid probe is provided with a target detection probe used for detecting target gene of pathogenic microorganism. The pathogenic microorganism comprises but not limits to vibrio cholerae, pathogenic escherichia coli, campylobacter jejuni, Yersinia enterocolitica, parahemolytic vibrio, salmonella, and shigella and Listeria monocytogenes. The invention also relates to a preparation method of the pathogenic microorganism DNA detection chip and provides applications of the pathogenic microorganism DNA detection chip in detection of pathogenic microorganism. Also a pathogenic microorganism DNA detection chip kit is provided.

The invention discloses a method for simultaneous and efficient amplification of CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells. The method comprises the steps as follows: the concentration of separated PBMC (peripheral blood mononuclear cells) is adjusted by a serum-free medium containing autologous plasma, an Anti-CD16 antibody, IL-2 and IL-15 are added, and then the mixture is transferred into a T175 culture flask for culture; an Anti-CD3 antibody and an Anti-CD137 antibody are added; a serum-free medium containing the autologous plasma, IL-2 and IL-15 is supplemented every two days according to the cell growth condition; the cell concentration is controlled to be about 1.5*10<6> / ml; and after culture is performed for 14-21 days, large quantities of high-purity CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells can be obtained simultaneously, and the total cell quantity can reach an effective value of the cell quantity required for adoptive cellular immunotherapy clinically for tumor. The method for simultaneous and efficient amplification of the CD<3+>CD<56+>CIK cells and the CD<3->CD<56+>NK cells is simple, convenient, effective and high in cell killing activity.

The invention discloses an isothermal amplification and detection technique based on CRISPR-chain substitution. An isothermal amplification kit based on CRISPR-chain substitution comprises a chain-substitution isothermal amplification reagent and further comprises wild-type Cas9 and / or nuclease defective Cas9 nickase, sgRNA pairs on the upstream and the downstream of a specific identification target DNA sequence, and an amplification primer pair of a chain-substitution isothermal amplification reaction, and the primer pair and a Cas9-sgRNA complex are complementary to a single-chain region exposed after being combined with target genes. The technique is easy to operate, high in anti-jamming ability, and accurate and reliable in tested result.

According to the invention, based on Illumina common tag library sequencing and methylation normal sequencing and combining a common library tag sequencing method, a new method for accurate detection of whole genome methylation sites by utilizing trace genome DNA (deoxyribonucleic acid) is built. According to the method, in the construction of a methylation library by using trace genome DNA, exogenous DNA is innovatively added to carry out bisulfite high-efficiency co-treatment; and at the same time, fragment size selection is not needed, and PCR (polymerase chain reaction) amplification is directly carried out after bisulfite treatment. By the method, the defects that in the common methylation sequencing, samples can not be mixed, the PCR amplification efficiency is low and the trace DNAsample can not be researched are overcome.

The invention relates to the technical field of gene detection, and provides a method and a primer for detecting mutation sites of all exon sequences of human BRCA1 and BRCA2 genes. The specific primers include forward and backward primers for amplifying all 24 exons covering the BRCA1 gene, and base sequences of the primers are as shown in SEQ ID NO: 001-062, as well as forward and backward primers for amplifying all 27 exons covering the BRCA2 gene, and base sequences of the primers are as shown in SEQ ID NO: 063-142. By virtue of a Sanger sequencing method, the method and the primer disclosed by the invention can be used for detecting the mutation of all exons of the BRCA1 gene and the BRCA2 gene and can be used for extending all exons of the entire BRCA1 and BRCA2 genes, covering all to-be-detected mutation sites. The method and the primer disclosed by the invention are quite high in specificity and accuracy, simple to operate and low in cost, and can be used for greatly enhancing the risk assessment of hereditary breast cancer and effectively reducing the onset risk of the breast cancer.

The invention discloses a method for amplification and activation of NK cells by K562 cells. The method comprises that through synergism of K562 cells transfected by transmembrane interleukin 21, CD14, CD19, CD86 and CD137, and low-concentration interleukin 2, NK cells are subjected to directed amplification and activation. Compared with the existing similar compounds, the compound provided by the invention has a stronger lymphocyte amplification and activation capability and higher efficiency. The method has wide prospects in immunological therapy.

With an appropriate signal transmission form, a drive pulse output circuit is mounted to an ink jet head, so that waveform distortion of an actuator drive pulse for jet of ink drops is inhibited or prevented. Modulated signal data in memory is read to output a modulated signal, which is power-amplified by a digital power amplifier. The amplified digital signal is smoothed to be output to an actuator as a drive pulse. As a result, the digital power amplifier having high amplification efficiency efficiently amplifies the power of the modulated signal, thereby eliminating the need to use a cooling unit. The drive pulse output circuit can be mounted to the ink jet heads, thereby shortening the transmission path of an actuator drive pulse, which inhibits or prevents any waveform distortion of the drive pulse. The transmission of the modulated signal data DATA is implemented with a low frequency.

The invention belongs to the technical field of nucleic acid detection, and relates to a mycobacterium tuberculosis complex detection kit and method based on a CRISPR-Cas12a system. According to the kit, the detection sensitivity is improved by adopting a recombinase polymerase amplification technology, CRISPR-Cas12a is used for specifically targeting a mycobacterium tuberculosis complex target sequence and then activating the bypass cutting activity of Cas12a, and a mycobacterium tuberculosis complex can be sensitively and specifically detected from sputum. The kit and method have the advantages of being non-invasive, capable of detecting the mycobacterium tuberculosis complex frequently and repeatedly, high in detection speed and the like. Compared with an existing PCR detection method,the method based on the CRISPR-Cas12a system does not need a thermal cycler with heating and cooling functions, can detect the mycobacterium tuberculosis complex in sputum through fluorescence reading, has the advantages of being low in cost, easy and convenient to operate, high in sensitivity, good in specificity and the like and is suitable for clinical large-scale application.

The invention discloses a nucleic-acid visual detection kit for a novel coronavirus SARS-CoV-2. The kit disclosed by the invention comprises a primer group and ingredients such as a RT-LAMP reaction solution, a positive control, a negative control and DEPC water. The primer group for detecting the SARS-CoV-2 comprises 6 primers in all, i.e., outer primers F3 and B3, inner primers FIP and BIP, andloop primers LF and LB and has a sequence shown in SEQ ID NO: 1 to NO: 6. The primer group and the kit can accurately detect an S gene of the SARS-CoV-2 and are free of a cross reaction with other coronaviruses and other viruses causing pneumonia. The SARS-CoV-2 can be detected in 15 minutes at the soonest through isothermal amplification at a temperature of 63 DEG C. The detection sensitivity reaches 1.6fg / microliter. The kit has the advantages of simplicity, convenience, rapidness, sensitiveness and specificity, a result can be directly decided through observing color changes of a reaction tube with naked eyes under natural light without expensive instruments, rapid and accurate detection on the SARS-CoV-2 can be achieved, and thus, the kit is applicable to on-site detection and clinicalcheckout in primary hospitals.

The invention provides an in vitro amplification method for efficient and highly cytotoxic natural killer (NK) cells. Novel artificial antigen presenting cells 4-1BBL-mIL-21-aAPC, such as 4-1BBL-mIL-21-K562 cells and the like, are constructed through stably expressing 4-1BB ligands (4-1BBL) and membrane immobilized interleukin 21 (mIL-21) on the surfaces of cell membranes, and by using the novel artificial antigen presenting cells as feeder cells for amplification, the NK cells are directly amplified from peripheral blood lymphocytes. Flow cytometry, cytotoxicity test and the like suggest that the amplified cells NK have high purity and strong cytotoxicity and have obvious killing effect on tumor cells.

The invention discloses an amplification primer for a grouper catostomidae mitochondrial genome complete sequence and a design method thereof, relates to groupers, and particularly relates to 16 pairs of amplification primers mainly used for specifically amplifying most grouper catostomidae mitochondrial genome complete sequences and a design method thereof. The invention provides an amplification primer for a grouper catostomidae mitochondrial genome complete sequence, a design method thereof and application thereof. The amplification primer for the grouper catostomidae mitochondrial genome complete sequence consists of 16 pairs of single-chain oligonucleotides, wherein each pair consists of one light-chain primer and one heavy-chain primer. The amplification primer for the grouper catostomidae mitochondrial genome complete sequence can be used in the aspects of variety authentication, geographical population authentication, germplasm resource evaluation and the like.

The invention discloses a multiplex LAMP detection primer, kit and method for six food-borne pathogenic bacteria in fruits and vegetables and belongs to the technical field of bacterial gene detection. A rapid detection primer set for the six pathogenic bacteria including listeria monocytogenes, enterobacter sakazakii, shigella spp, staphylococcus aureus, salmonella spp and escherichia coli O157:H7 is designed, and multiplex LAMP reaction is performed on the genome DNA of the bacteria extracted from a sample to be detected in the same reaction system by use of the detection kit including the primer set to determine whether the sample contains the six food-borne pathogenic bacteria or not. The multiplex LAMP detection primer is high in specificity and sensitivity and can accurately detect the genome DNA of the six food-borne pathogenic bacteria in the same reaction system, can realize simple and convenient, quick and accurate detection, is suitable for on-site rapid detection and has significance on improving the pathogenic bacterium analysis and detection technology and the fruit and vegetable edible quality security.

The invention relates to a CRISPR detection primer set for MTBC (Mycobacterium Tuberculosis Complex) and use thereof, and belongs to the technical field of gene detection. The primer set includes an amplification primer pair and a crRNA; the amplification primer pair is used to amplify a common conserved sequence of M. tuberculosis, M. bovis, M. africanum, M. canettii, and M. microti; the crRNA includes an anchor sequence and a guide sequence, wherein the anchor sequence is bound to a cas protein, and the guide sequence is matched with a targeted RNA fragment in the common conserved sequence.Through the primer set based on CRISPR detection technology, the detection of common conserved sequences of M. tuberculosis, M. bovis, M. africanum, M. canettii, and M. microti by CRISPR detection canquickly perform on-site detection of MTBC, and have the advantages of high specificity, high sensitivity and simplicity.

The invention discloses a method for detecting nucleic acid or cells based on enzymatic cycle amplification and nano-particle reinforced SPR (surface plasmon resonance). Strand displacement cycle amplification and rolling circle replication amplification are triggered by a target DNA (deoxyribonucleic acid), a double-signal amplification reaction is formed, a magnetic bead is taken as a DNA carrier and serves as a transfer station for signal amplification, a magnetic bead bio-barcode probe has a strand displacement cycle amplification reaction as a reaction unit and also increases the load capacity of a rolling circle replication product as a signal amplification carrier, a multi-stage signal amplification system is formed, high-sensitivity detection of the target DNA is realized with gold nano-particles as signal probes with an SPR technique, and the limit of detection can reach 1fM.

The invention discloses a gene engineering cell and application thereof in NK (Nature Killer) cell proliferation, and provides an in vitro proliferation method of an NK cell with high efficiency and high cytotoxicity. A 4-1BB ligand (4-1BBL) and a membrane fixed interleukin 21 (mbIl-21) are stably expressed on the surface of a cell membrane by adopting a genetic engineering technology for constructing 4-1BBL-mbIl-21-gene engineering cells (4-1BBL-mbIl-21-Gene Engineering Cells, 4-1BBL-mbIl-21-GEC), such as 4-1BBL-mbIl-21-K562 cells, to be used as feeder cells for proliferation, and the NK cells are directly proliferated from peripheral blood monouclear cells, thus the operation is simpler and easier. Studies such as cell counting, flow cytometry and cytotoxicity tests indicate that high cell proliferation times, high NK cell purity, strong cytotoxicity and remarkable cancer cell killer effect are achieved.

The invention relates to a double-Z operation type solid laser batten amplifying device. The double-Z operation type solid laser batten amplifying device comprises batten laser media, a semiconductor pump source, a beam shaping system, endoscopes and a cooling device, wherein pumping lasers generated by the semiconductor pump source serve as a seed laser system through the beam shaping, enter from one end face of a batten, are transmitted forward in a total internal reflection mode between two large surfaces of the batten and output from the other end face, are reflected after reaching the endoscope on the right side, enter from the end face of the batten, are transmitted forward in a total internal reflection mode between the two large surfaces of the batten, are reflected after reaching the endoscope on the left side, enter the batten again, and finally are output from the endoscope on the right side after being reflected repeatedly. According to the double-Z operation type solid laser batten amplifying device, bidirectional Z type multipass transmission of the seed lasers along the Y axis and the Z axis in the batten is achieved, amplification with high power and high efficiency is achieved; sodium beacon laser amplification is high in power, high in beam quality, and high in efficiency, and the double-Z operation type solid laser batten amplifying device can be used for generating sodium beacon lasers which are high in power, high in beam quality, and small in line width.

The invention provides a high-contrast femtosecond laser generating device comprising a first optical parallel moving table and a first optical parameter crystal. The first optical parallel moving table is suitable for controlling the optical path difference between a first signal light and a first pump light, so as to achieve the time synchronization of the first signal light and the first pump light; and the first optical parameter crystal is suitable for receiving the first signal light and the first pump light which are in time synchronization, carrying out optical parameter enlargement, and outputting a first enlarged signal light. The high-contrast femtosecond laser generating device further comprises a second optical parallel moving table and a second optical parameter crystal, the second optical parallel moving table is suitable for controlling the optical path difference between the first enlarged signal light and the second pump light, so as to achieve the time synchronization of the first enlarged signal light and the second pump light; the second optical parameter crystal is suitable for receiving the first enlarged signal light and the second pump light which are in time synchronization, carrying out optical parameter enlargement, and outputting a second enlarged signal light. The generating device provided by the invention can improve the contrast and the optical parameter enlargement efficiency of the femtosecond laser and increase parameter bandwidth in the process of the optical parameter enlargement.

The present invention discloses an autosome STR gene locus fluorescent labeling composite amplification kit having enhanced identification ability, wherein 27 STR gene loci can be simultaneously amplified with the kit and comprise 24 autosome STR gene loci such as D3S1358, TH01, D21S11, D18S51, Penta E, D12S391, D6S1043, D2S1338, D1S1656, D2S441, D5S818, D13S317, D7S820, D19S433, CSF1PO, Penta D, vWA, D8S1179, TPOX, FGA, D16S539, D22S1045, SE33 and D10S1248, 2 Y chromosome gene loci such as Y-indel and DYS391, and a sex-determining locus Amel. According to the present invention, with the application of the kit to perform the DNA gene detection, all the gene loci of the current mainstream products at home and abroad are contained, the currently existing DNA database in most countries are covered, the compatibility problem is not required to be worried about, the accumulation individual recognition rate and the combined paternity non-exclusion probability of the system are improved, and the individual discriminability is improved on the whole.

The invention, blonging to plant biotechnology field, relates a method of checking the virus of B hepatitis genovariation. The invention utilize the fluorescent light definite quantity PCR MGB Taqman probe technique, according to the YMDD nucleic acid sequence of B hepatitis virus, designing the genovariation of primer and probe checking HBV YMDD region. The invention adopts PCR reaction mixture A, B, Tap enzymes, HBV DNA extractant, and prepares diagnosis agent case after optimization and improvement with ddH20. The method can rapidly and exactly check the gene mutation situation of privileged site, quantitate the ratio of hybrid mutational virogene, quckly check the clinical hepatitis B drug therapia drug tolerance mutation. The method and agent case are not confined by medical equipment, so it is very easy to popularize.

The invention belongs to the field of cell engineering, in particular to a method for high-efficient expansion and cryopreservation of NK (Natural Killer) cells and an application of the method. According to the invention, fusion genes are subjected to gene recombination to an AAVS1 (Adeno-Associated Virus Integration Site 1) transposon through a plasmid, and the freezing / thawing step is introduced to the NK cell expansion step taking a modified K562 cell line as a basis in the intermediate stage of cell expansion; finally, the NK cells are expanded and saved. A K562-mbIL15-41BBL-mbIL21 cell line developed by the integration technology of site specific genes is used, and the activity, purity and cytotoxicity of the NK cells are high to realize high expansion efficiency of the NK cells to improve NK expansion. The method for the high-efficient expansion and cryopreservation of the NK (Natural Killer) cells and the application of the method for the high-efficient expansion and cryopreservation of the NK (Natural Killer) cells disclosed by the invention have great market prospects and economic values.

The invention relates to a DNA methylation biomarker composition for bladder cancer detection. The biomarker composition comprises any two or more of sequences shown in the formulas of SEQ ID NO.1 toSEQ ID NO.22, or any two or more of completely complementary sequences shown in the formulas of SEQ ID NO.1 to SEQ ID NO.22. The invention also relates to a detection kit for the methylation biomarkercomposition. The bladder cancer occurrence is discriminated and analyzed by using the composition in the co-methylation state of a plurality of specific methylation regions, the specific methylationcomposition has high sensitivity for discriminating the bladder cancer occurrence, and the detection method is simple, convenient and feasible. The primer pair composition of the kit overcomes the defect of false positive results caused by single methylation site detection mismatch in primer sequence design, and considers the interaction between multiple methylation biomarker primer and probe paircomposition.

The invention discloses a kit for mesenchymal stem cell culture and its application. The kit provided by the invention includes a cell culture solution A and a cell culture solution B. The cell culture solution A is a Knockout-DMEM culture medium containing 80-120ng / ml cytokine LIF and 80-120ng / ml cytokine bFGF. The cell culture solution B is fetal bovine serum. The kit provided by the invention can separate umbilical cord mesenchymal stem cells from an in-vitro umbilical cord and culture the umbilical cord mesenchymal stem cells, and has the advantages of rapid cell expansion speed and high expansion efficiency. After multiple subculturing, cell totipotency can still be maintained. In the invention, the problems of umbilical cord mesenchymal stem cell separation purity, quantity and primary culture time are solved, and a lot of umbilical cord mesenchymal stem cells can be obtained. Thus, the kit provided in the invention provides abundant sources for making use of umbilical cord mesenchymal stem cells to conduct further experimental research.

The invention relates to a method for expanding activated lymphocyte, comprising the steps of allowing biologic sample containing lymphocytes to contact immobilized anti-CD 3 antibodies, then cultivating activated lymphocytes with culture mediums in different IL-2 concentrations to expand the activated lymphocytes. The invention also relates to a cultivation system used for expanding activated lymphocytes. The expanded activated lymphocytes can be applied in the immunotherapy of tumor, infectious diseases, and congenital or acquired immunodeficiency diseases.

The invention provides a high-contrast femtosecond laser pulse generation device which comprises a first optical parameter crystal which is used for receiving first signal light and first pump light, amplifying an optical parameter and outputting first idler frequency light. Selectively, the high-contrast femtosecond laser pulse generation device further comprises a second optical parameter crystal which is used for receiving second signal light and second pump light, amplifying an optical parameter and outputting second idler frequency light. The second signal light is the first idler frequency light. Selectively, the high-contrast femtosecond laser pulse generation device further comprises a third optical parameter crystal which is used for receiving third signal light and third pump light, amplifying an optical parameter and outputting third idler frequency light. The third signal light is the second idler frequency light. On one hand, the contrast of a femtosecond laser and amplifying efficiency of the optical parameter are improved, and on the other hand, the amplified parameter bandwidth in the optical parameter is also increased.

The invention discloses a small-molecular compound combination for differentiated cell reprogramming, and a reagent kit and an application of the small-molecular compound combination. The small-molecular compound combination includes the following components: a TGF-beta inhibitor, a WNT / beta-catenin agonist, a cAMP agonist and a PKC inhibitor; furthermore, the small-molecular compound combinationalso includes at least one of an RAR agonist, a DNMT inhibitor, an HMT inhibitor, a histone demethylase inhibitor, ascorbic acid, a JNK inhibitor, an ROCK inhibitor and a lysine deacetylase inhibitor.Differentiated cells can be reprogrammed into mesenchymal stem cells by phased induction with the small-molecular compound combination, all steps can be subjected to precise quality control, and standardization operation and large-scale production are convenient. The method provided by the invention has wide donor sources, a patient himself can be used as a donor, and the mesenchymal stem cells needed for basic research, clinical treatment or tissue engineering production can be obtained in relatively short time.

The invention discloses a pre-amplification method for trace DNA applied in medicolegal expertise. The method aims at amplifying the trace DNA to obtain high-concentricity DNA template so as to facilitate following medicolegal expertise. The method mainly comprises the steps of exploring the optimum concentration and proportion volume of combined primers; and the amplification conditions are further optimized, the trace DNA aims for being amplified with high efficiency and high fidelity, so that the possibility for the next legal medical expert is provided for determining individuals.