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195 results about "Lysis buffer" patented technology

A lysis buffer is a buffer solution used for the purpose of breaking open cells for use in molecular biology experiments that analyze the labile macromolecules of the cells (e.g. western blot for protein, or for DNA extraction). Most lysis buffers contain buffering salts (e.g. Tris-HCl) and ionic salts (e.g. NaCl) to regulate the pH and osmolarity of the lysate. Sometimes detergents (such as Triton X-100 or SDS) are added to break up membrane structures. For lysis buffers targeted at protein extraction, protease inhibitors are often included, and in difficult cases may be almost required. Lysis buffers can be used on both animal and plant tissue cells.

Cell lysis buffer and protein pretreatment method based on digital microfluidic system

The invention relates to a cell lysis solution, which is prepared from the following active components: 0.08 to 0.2 percent of DDM (Dichloromethane), 35 to 45 mM of CAA (Ceamic Acid Acetic Acid), 5 to 15 mM of TCEP (Trichloroethylene Phosphate), 85 to 100 mM of TEAB (Tetraethyl Ammonium Bromide), 0 to 0.25 percent of Protease MAX and 0 to 0.25 percent of RapiGest SF (SF). The invention further provides a single-cell protein pretreatment method based on the digital microfluidic system, the cell lysis solution is used for lysis, the number of detectable proteins and the number of detectable peptide fragments can be high, and the method can be well used for single-cell proteomics analysis.
Owner:GUANGDONG ACXEL MICRO & NANO TECH CO LTD

Systems, methods, and devices for pathogen identification

Described herein are systems, methods, and devices for pathogen identification. A system includes a housing configured to receive a sample comprising pathogen(s), a pipettor system disposed inside the housing, centrifuge(s) disposed inside the housing, a mechanical agitator disposed inside the housing, and a controller. The controller is configured to transfer the sample to a processing tube using the pipettor system, centrifuge the processing tube using the centrifuge(s) to concentrate the pathogen(s) in the sample, remove a fluid from the processing tube using the pipettor system, leaving the concentrated pathogens in the processing tube, add a lysis buffer to the processing tube using the pipettor system, move the processing tube to the mechanical agitator using the pipettor system, and agitate the processing tube using the mechanical agitator to perform lysis of the concentrated pathogens. The system is further configured to perform PCR using a nucleic acid extracted from the sample.
Owner:DEEPULL DIAGNOSTICS SL

Preparation method and application of gynostemma pentaphylla exosome

The invention discloses a preparation method and application of a gynostemma pentaphylla exosome. According to the method, a high-activity callus is obtained by optimizing a callus induction culture medium, and a suspension culture system is established to realize biomass accumulation with the dry cell weight of 19.8 + / -0.7 g / L. A sequential progressive composite induction strategy (methyl jasmonate is added after chitosan is pre-induced for 24 hours) is innovatively adopted, so that the saponin content is remarkably increased to 102.6 + / -4.8 mg / g DW. Exosomes (P-Exos) are extracted from induced cells through an enzymolysis-magnetic bead affinity purification technology (WGA modified magnetic bead affinity capture after mild enzymolysis of a cracking buffer solution), the yield reaches 1.45 + / -0.08 mg / g cells, and the extraction efficiency is remarkably improved compared with that of a traditional gradient centrifugation method (C-Exos). Moreover, the gynostemma pentaphyllum exosome prepared by the invention can effectively improve the synthesis of skin collagen, has a better effect, can be applied to medicines for promoting skin wound repair or can be added into anti-aging cosmetics as a functional component, and has a very high application value.
Owner:NANJING BAISIHE BIOTECHNOLOGY CO LTD

Gelatin microcarrier lysate as well as preparation method and application thereof

The invention discloses a gelatin microcarrier lysate as well as a preparation method and application thereof. The gelatin microcarrier lysis buffer comprises lyase, a cell protective agent, a cell energy metabolism protective agent, an antioxidant, a buffer solution and a chelating agent. The gelatin microcarrier lysate has extremely strong protectiveness to cells, abandons damaging trypsin and EDTA, and innovatively adds a cell protection combination of a cell protective agent L-arginine, a cell energy metabolism protective agent sodium pyruvate and an antioxidant vitamin C, so that the cell survival rate (gt; 98%) and the retention rate (gt; the method is high in cell yield, good in safety and wide in range.
Owner:SUZHOU HUACHEN BIOTECHNOLOGY CO LTD

Rapid culture system and culture method of mycoplasma pneumoniae

The invention discloses a rapid culture system of mycoplasma pneumoniae. The rapid culture system comprises a growth-promoting additive A, a growth-promoting additive B and a growth-promoting additive C, wherein the growth-promoting additive A is A549 cell lysis buffer; the growth promoting additive B is a mixture of deoxyribonucleoside triphosphate and / or ribonucleoside triphosphate. The rapid culture system can promote rapid growth and reproduction of the mycoplasma pneumoniae, fully meets the nutritional requirements and growth requirements of the mycoplasma pneumoniae, and is of great significance to isolated culture, enrichment culture, epidemic disease prevention and control and molecular diagnosis of the mycoplasma pneumoniae. The invention also provides a culture method of the mycoplasma pneumoniae. The invention also provides application of the rapid culture system of the mycoplasma pneumoniae in in-vitro cell culture of the mycoplasma pneumoniae.
Owner:SOUTHWEST MEDICAL UNIV

Method and system for collaborative analysis of molecular interaction state based on multi-mode optical parameters of rare earth up-conversion nano material

The invention belongs to the technical field of biomolecular interaction detection, and particularly relates to a method for synergistically analyzing a molecular interaction state based on multimodal optical parameters of a rare earth up-conversion nano material. The method comprises the following steps: S1, providing a ligand coated rare earth up-conversion nano material UCNPs probe; s2, enabling the rare earth up-conversion nano material UCNPs probe to be in contact with a solution sample to be detected; s3, exciting the rare earth up-conversion nano material UCNPs probe by using near-infrared pulse laser, and collecting the following optical parameters: luminous intensity change, molecular layer thickness / mass change delta D, luminous life change delta tau and emission peak displacement delta lambda; and S4, based on at least two of the parameters, cooperatively analyzing and judging the interaction state of the ligand on the surface of the probe and the molecules in the solution sample to be detected. The molecular interaction state is synergistically analyzed based on changes of multiple optical parameters, and the technical problem that an existing molecular interaction analysis technology cannot be applied to complex biological samples (serum and cell lysis buffer) is solved.
Owner:SHANGHAI LEIMENGKE TECHNOLOGIES CO LTD

CfDNA extraction kit and extraction method

The invention provides a cfDNA extraction kit and an extraction method. The cfDNA extraction kit provided by the invention comprises protease K, a lysis buffer solution, magnetic beads and a binding buffer solution, by limiting the composition of the lysis buffer solution and the binding buffer solution, not only can the extraction efficiency of cfDNA be effectively improved, but also the purity of the cfDNA can be remarkably improved, the cfDNA of 100-200 bp can be effectively enriched, and the kit is convenient in component acquisition, low in cost and suitable for industrial production. Meanwhile, the extraction operation is simple and suitable for high-throughput treatment.
Owner:SHANGHAI JINFUKANG PHARMACEUTICAL ENGINEERING TECHNOLOGY CO LTD

Nucleic Acid Extraction Kit for Fecal Samples, Preparation Method and Extraction Method

The present invention provides a nucleic acid extraction kit, a preparation method and an extraction method for fecal samples, which relates to the field of biotechnology. The kit includes: proteinase K, primary lysis buffer and lysis-binding buffer; the concentration range of proteinase K is 10-30 mg / mL; the primary lysis buffer contains: 50-200 mM Tris-HCl with pH 6.0-8.5, 0.5-30% (w / v) SDS, 1-10 mM EDTA with pH 8.0-9.0; the lysis-binding buffer contains: 50-200 mM Tris-HCl, 1-15 M guanidine hydrochloride, 1-10 mM EDTA, 0.1-5% Triton X-100, 0.1-5% Tween-20, with pH 7.0-8.0. The DNA nucleic acid extracted and purified by the kit has a high yield and good purity, and solves the problem of short storage time of the DNA extracted from fecal samples.
Owner:JIANGSU COWIN BIOTECH CO LTD +2

Preparation method of thermostable T7 RNA polymerase and thermostable T7 RNA polymerase

The invention provides a preparation method of thermal-stability T7RNA polymerase and the thermal-stability T7RNA polymerase, belongs to the field of biology, and can solve the problems that an existing T7RNA polymerase preparation process is complex, the expression culture period is long, multi-step purification treatment is needed, liquid change treatment needs to be carried out independently before storage, and the production cost is high. The method comprises the following steps: transferring plasmids containing a T7RNA polymerase gene sequence into expression host bacteria M15, culturing, screening, cloning, adding glycerol for strain preservation, and establishing a strain three-level library; the method comprises the following steps: taking glycerol bacteria, carrying out streak resuscitation on a flat plate, selecting monoclone, inoculating the monoclone into an LB culture medium, culturing, carrying out enlarged culture, carrying out IPTG induced expression, centrifugally collecting thalli, cleaning, adding a lysis buffer solution, resuspending, carrying out ultrasonic cell disruption, centrifuging, and collecting supernate; purifying by nickel column affinity chromatography, removing impure protein by an impure washing buffer solution, eluting target protein, purifying by matching with anions and cations, and collecting the purified protein; and mixing the purified protein with a storage buffer solution, and subpackaging and storing. The method can be applied to rapid preparation of the thermal-stability T7RNA polymerase.
Owner:SHANDONG SIKEJIE BIOTECHNOLOGY CO LTD

Preparation method and application of red blood cell lysis buffer for Rh double-group typing

The invention provides a preparation method and application of a red blood cell lysis buffer for Rh double-group typing. The preparation method comprises the following steps: (1) preparing a stock solution 1; (2) preparing a stock solution 2; (3) mixing the stock solution 1 with H2O to obtain an application solution 1; (4) preparing application liquid 2; (5) adding 50 [mu] L of to-be-detected sample near-albuginea layer hematopoietic cells into 1 mL of the application liquid 1, and uniformly mixing; (6) centrifuging the mixed solution obtained in the step (5), and discarding the supernate; washing with normal saline under the condition of 1000g each time, and centrifuging; discarding supernate, dripping 2-3 drops of normal saline, and uniformly mixing; and (7) adding one drop of erythrocyte suspension and one drop of corresponding antibody into the blank card, and centrifugally observing the result. According to the preparation method of the red blood cell lysis buffer for Rh double-group typing, the detection rate is high, the preparation method is simple, the cost is low, and the red blood cell lysis buffer is very suitable for being popularized and popularized in primary medical institutions.
Owner:SHENZHEN UNIV GENERAL HOSPITAL

Extraction method of genome DNA of plant rich in secondary metabolites and buffer solution

The invention relates to a method for extracting genome DNA of plants rich in secondary metabolites and a buffer solution, and belongs to the technical field of molecular biology and botany. The method solves the technical problems of low DNA extraction efficiency, poor purity, easy degradation and the like when a traditional DNA extraction method is used for treating plant tissues rich in secondary metabolites such as alkaloid, polyphenol and the like. Comprising the following steps: adding a complexing agent such as polyethylene glycol or polyvinylpyrrolidone when grinding plant tissues in a liquid nitrogen environment; splitting by using a cell wall splitting buffer solution containing dithiothreitol and a nonionic surfactant; carrying out DNA release and extraction at 60-70 DEG C by adopting a CTAB (Cetyltrimethyl Ammonium Bromide) extraction buffer solution containing polyethylene glycol and papain; and then purifying and precipitating to obtain high-purity genome DNA (Deoxyribose Nucleic Acid). The method can effectively remove alkaloid, protein and other impurities, significantly improves the DNA yield and purity, and is suitable for genome sequencing, genetic resource protection, medicinal plant molecular identification and the like of plants with high secondary metabolites such as Stephania kwangsiensis and the like.
Owner:广西农业职业技术大学

Brevundimonas sp. Fermentation medium and application thereof in prevention and treatment of plant root-knot nematodes

The invention discloses a brevundimonas sp. Fermentation medium and application thereof in prevention and treatment of plant root-knot nematodes. Each liter of the brevundimonas chinensis fermentation medium comprises the following components by mass or volume: 6-10 g / L of yeast powder, 16-18 g / L of peptone, 31-45 ml / L of nematode lysis buffer, 0.8-2.5 g / L of chitin and 5.6-6 g / L of yeast extract, and the pH value of the brevundimonas chinensis fermentation medium is 6.5-6.8. The application of the fermentation broth culture medium refers to the application of the obtained fermentation broth in prevention and treatment of plant root-knot nematodes. According to the present invention, with the fermentation medium, the fatality rate of the obtained fermentation broth on the plant root-knot nematode exceeds 95%, the action time does not exceed 12 h, the nematode can be rapidly killed before the larva enters the plant, and the good application prospect is provided in the plant root-knot nematode prevention and control.
Owner:YUNNAN UNIV

Liriodendron tulipifera cell nucleus extraction method suitable for CUTTag technology and application of liriodendron tulipifera cell nucleus extraction method

The invention discloses a method and a device suitable for CUAMP. The invention discloses a method for extracting liriodendron tulipifera cell nucleuses by a Tag technology and application of the liriodendron tulipifera cell nucleuses. Hybridized liriodendron tulipifera calluses are used as materials, and protoplasts are released through enzymolysis solution vacuumizing and mild enzymolysis for 1.5 h; the method comprises the following steps: sequentially purifying by using a W5 solution and mannitol, detecting the activity by FDA, cracking by using an NE buffer solution, and washing by using a Wash buffer solution in two steps, thereby obtaining 2 * 10 < 6 >-3 * 10 < 6 > high-purity and high-integrity cell nucleuses. The obtained core is clean in background and complete in membrane structure, and can be directly used for CUTamp; tag is used for building a library, so that the magnetic bead capturing efficiency is improved by 46%, and protein-DNA interaction high-throughput sequencing under the condition of low sample size is realized. The method disclosed by the invention is simple and convenient to operate and good in repeatability, and provides key technical support for epigenetic research of rare tree species such as liriodendron tulipifera.
Owner:NANJING UNIV +1

Epidermal fibroin (FLG) gene and protein detection method based on oral cell non-invasive sampling

The invention discloses an FLG gene and protein detection method based on oral cell non-invasive sampling, which comprises the following steps: Step 1: sample collection: collecting oral cells through mouth rinsing of a subject; step2: RNA extraction and gene expression analysis: extracting oral cell total RNA in the oral cells in the step 1 by using a Trizol reagent, and analyzing FLG gene expression through qRT-PCR (Quantitative Reverse Transcription-Polymerase Chain Reaction); 3, protein extraction and immunoblotting analysis are conducted, specifically, protein is extracted from the oral cells in the step 1 through an SDS lysis buffer solution, and FLG protein expression is detected through the Western blot technology. According to the present invention, the subject rinses the mouth and collects the oral cavity cells, such that the convenient, non-invasive and efficient sample collection is achieved.
Owner:WENZHOU-KEAN UNIV

Method for accurately identifying and screening morchella strain by using molecular marker

The invention discloses a method for accurately identifying and screening morchella strains by using molecular markers, and belongs to the technical field of edible mushrooms. The method comprises the following steps: transferring a purified morchella strain to a culture medium for dark light cultivation; adding the cultured morchella hyphae into liquid nitrogen, grinding into powder, then adding a lysis solution, and carrying out lysis; adding modified magnetic beads into the lysate for adsorption, and then filtering, washing and eluting to obtain a DNA extracting solution; carrying out PCR (Polymerase Chain Reaction) amplification on the DNA; then carrying out high-resolution detection on an ISSR product; and then, carrying out functional SNP-KASP typing. According to the method, by fusing ISSR polymorphism analysis and a functional SNP-KASP typing technology, magnetic beads are used for adsorbing DNA to remove polysaccharide interference, and on the basis that the low cost advantage of traditional ISSR is reserved, three functional markers are completed, and accurate screening of yield, heat resistance and flavor is achieved.
Owner:HEILONGJIANG ACADEMY OF SCI

A cfDNA extraction kit and extraction method

The present invention provides a cfDNA extraction kit and method. The cfDNA extraction kit provided by the present invention includes proteinase K, a lysis buffer, magnetic microbeads, and a binding buffer. By limiting the composition of the lysis buffer and the binding buffer, it can not only effectively improve the extraction efficiency of cfDNA, but also significantly increase the purity of cfDNA and effectively enrich cfDNA of 100-200 bp. In addition, the kit components are easily available and low-cost, and the extraction operation is simple and suitable for high-throughput processing.
Owner:SHANGHAI JINFUKANG PHARMACEUTICAL ENGINEERING TECHNOLOGY CO LTD

Genetic engineering strain and application thereof in D2P

The invention provides a gene engineering strain and application thereof in D2P, and particularly, the gene engineering strain is obtained by modifying a gene of a Ubp family originally existing in eukaryotic cells, the expression or activity of Ubp in the gene engineering strain is reduced, the IVTT activity of a strain lysis buffer can be effectively improved, and the D2P production capacity is remarkably improved.
Owner:KANGMA (SHANGHAI) BIOTECH LTD

Virus-like vector lysis buffer and virus-like vector lysis method thereof

The invention provides a virus-like vector lysis solution and a virus-like vector lysis method. The virus-like vector lysis solution comprises Tris, EDTA (Ethylene Diamine Tetraacetic Acid), NaCl, TritonX-100, sodium deoxycholate, SDS (Sodium Dodecyl Sulfate), Tween 20 and a protease inhibitor mixture. Cas9 protein is wrapped in the viroid vector; the acquisition method of the viroid vector comprises the step of transfecting plasmids containing a structural gene sequence for expressing the viroid vector into virus producing cells to obtain the viroid vector. The cracking method of the viroid vector comprises the following steps: S1, using: carrying out cracking treatment on a viroid vector solution by using the cracking solution; and S2, performing Cas9 protein content detection on substances released after the carrier is cracked. The invention establishes a formula of the special lysate for VLP and a corresponding cracking method, and ensures high cracking efficiency.
Owner:SHANGHAI BDGENE TECH CO LTD

Cell lysis buffer clarification and separation device for plasmid production

The utility model provides a cell lysis solution clarification and separation device for plasmid production. The cell lysis solution clarification and separation device comprises a phase separation container, at least one air inlet valve arranged at the bottom of the phase separation container, and at least one drain valve arranged on the side wall of the phase separation container, the phase separation container is used for standing and layering the cell lysis solution; the air inlet valve is used for blowing air to the bottom of the phase separation container; and the drain valve is used for discharging the clarified layer of the cell lysis solution after standing and layering. The clarification and separation device for the cell lysis solution is particularly suitable for large-scale production of plasmids, the purification cost is reduced while the separation efficiency is improved, RNA residues are effectively separated out, and large-scale popularization and application are facilitated.
Owner:SHANGHAI HUIHENG GONGDAO BIOTECHNOLOGY CO LTD

Nitrate reduction oceanobacter sp. And application thereof

The invention relates to nitrate reduction pelagic bacteria and application thereof. The nitrate reductive ocean bacillus can be fermented to produce pyrroloquinoline quinone (PQQ) and trichothecenes toxin degrading enzyme, the trichothecenes toxin degrading enzyme mainly exists in bacterial cell lysis buffer, and the degradation rate of the trichothecenes toxin degrading enzyme to vomitoxin is 83%. The bacterial strain fermentation liquor can be used for degrading 39.3 percent of nivalenol and 100 percent of 15-acetyl deoxynivalenol. The strain or the trichothecene toxin degrading enzyme produced by the strain can be used for complete feed, concentrated feed, premixed feed, feed raw materials such as grain, corn, peanut meal, wheat and oat, silage, grain, grain and oil, grain processing and agricultural and sideline products, aged grain, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour, corn flour and corn flour. Through biodegradation of industrial ethanol processing byproducts and trichothecenes in food, the quality safety of feed and food can be improved.
Owner:CHINA AGRI UNIV

A method for extracting DNA from kappaphycus alvarezii

PendingCN122750672ATE bufferLysis buffer
A method for extracting DNA from Kappa longicornis, relating to the field of molecular biology, includes the following steps: grinding a Kappa longicornis sample into powder; adding lysis buffer preheated to 60–65°C to the powder and incubating at 60–65°C; adding Proteinase K for protein digestion; adding RNase A for RNA digestion; extracting with a mixture of chloroform and isoamyl alcohol, centrifuging, and collecting the supernatant; adding pre-cooled isopropanol to the supernatant for precipitation, centrifuging, and obtaining the DNA precipitate; washing and drying the DNA precipitate; and dissolving the DNA in TE buffer or nuclease-free water. The beneficial effects of this invention are as follows: by optimizing the lysis buffer and combining it with stepwise enzymatic hydrolysis and organic extraction processes, polysaccharides and pigments of *Kappa longicornis* can be efficiently removed, and the extracted DNA concentration is >300 ng / μL, with a purity of A260 / A280≈1.76 and A260 / A230≈2.21, meeting the requirements of downstream molecular experiments; at the same time, reduced glutathione is used instead of 2-mercaptoethanol, making the operation safe and odorless, with low reagent costs, and suitable for batch sample processing.
Owner:SHANTOU UNIV

ODC1 enzyme activity detection method based on CB6 probe

The invention relates to a method for determining enzyme activity of ODC1 (ornithine decarboxylase 1), which comprises the following steps: digesting, centrifuging and resuspending cells, and taking a part of sample to detect protein concentration; the method comprises the following steps: cracking cells by using a specially-made enzyme activity cell lysis solution and a protein lysis solution, and ensuring full cracking through ultrasonication; carrying out quantitative analysis on the protein in the lysate by adopting a BCA kit; in the enzyme activity detection stage, a cell lysis solution and an enzyme activity detection working solution containing a specific fluorescent probe (CB6) are mixed and react at 37 DEG C for a certain time, and then the change of fluorescence intensity is determined by a multifunctional fluorescence microplate reader, so that the enzyme activity of the ODC1 is evaluated. According to the method, accurate determination of the ODC1 enzyme activity is realized by optimizing cell treatment, protein quantification and enzyme activity detection processes. The method has the advantages of simplicity and convenience in operation, high sensitivity, good specificity and the like, and is suitable for the fields of basic research, drug screening and the like related to the ODC1 enzyme activity.
Owner:NINGXIA UNIVERSITY

Pathogenic microorganism nucleic acid co-extraction kit, extraction method and application

The invention belongs to the technical field of molecular biology, and particularly relates to a pathogenic microorganism nucleic acid co-extraction kit, an extraction method and application. The invention provides a lysis solution for nucleic acid co-extraction of pathogenic microorganisms. The lysis solution comprises a first guanidine salt, an ionic surfactant, a first sodium salt and a first nonionic surfactant. According to the splitting system, guanidine salt is adopted and matched with a certain content of sodium salt and a surfactant, a sample is split and precipitated under the grinding condition, nucleic acid cannot be damaged in the crushing process, and pathogenic microorganism DNA and RNA with high concentration and complete fragments can be extracted.
Owner:WUHAN MGI TECH CO LTD

Kidney cancer detection method based on BAFF / TACI interaction molecule and kit thereof

The invention discloses a kidney cancer detection method based on BAFF / TACI interaction molecules and a kit thereof, and the method comprises the following steps: S1, sample pretreatment: collecting a serum or tissue supernatant sample, carrying out centrifugal separation on the serum for 10 min under the conditions of 3000 rpm and 4 DEG C, adding an RIPA lysate into the tissue sample for homogenization, then carrying out centrifugation for 15 min under the conditions of 12000 rpm and 4 DEG C, and taking the supernatant; diluting the sample with PBS (phosphate buffer solution) containing 0.1% of BSA (bovine serum albumin); s2, constant-temperature three-channel ELISA detection: adopting a pre-coated three-channel 96-hole detachable elisa plate (7), and synchronously detecting in a constant-temperature environment of 37 + / -0.5 DEG C; S3, result judgment: the critical values are sBAFF > = 85pg / mL, sTACI > = 42pg / mL, and BAFF-TACI complex > = 12pg / mL, and if any condition is met, the kidney cancer is positive, and the kit has the advantages that the kit is suitable for early screening, clinical diagnosis, pathological staging and prognosis evaluation of the kidney cancer, and has a good application prospect. Through constant temperature control and three-channel independent hole design, synchronous and accurate detection of BAFF, TACI and BAFF-TACI compounds in various samples such as serum, plasma, tumor tissue and the like is realized.
Owner:WUXI PEOPLES HOSPITAL

Ketamine small molecule probe, and preparation method therefor and use thereof

Disclosed in the present invention are a ketamine small molecule probe, and a preparation method therefor and the use thereof. Two small molecular probes are designed and synthesized, and the blank in the prior art is filled. A target fishing experiment is performed on the total protein in a cell lysis buffer by means of using the two probes, and 61 and 70 target proteins are obtained, respectively. In addition, more than three technical repetitions verify that the two probes can stably bind to the target proteins. A screened SIRT2 protein is used as a target protein of ketamine, and a reference basis is provided for the synthesis and preparation of an anti-inflammatory related drug.
Owner:CHANGCHUN INSTITUTE OF APPLIED CHEMISTRY CHINESE ACADEMY OF SCIENCES

Method for extracting cell nucleus from animal tissue

The invention discloses a method for extracting cell nucleuses from animal tissues, and relates to the technical field of biology. The background technology indicates that cell nucleus extraction is crucial to multidisciplinary research, but a traditional method has the problems of complex operation, time consumption, low purity, easiness in damaging cell nucleuses and the like, and the prior art is still limited although the prior art is improved. The method comprises the following steps: preparing a sample, and taking fresh or cryopreserved animal tissues; homogenizing the tissues, namely putting the tissues into a homogenizer, and homogenizing at a low speed by adding a cracking buffer solution; carrying out cell lysis, and incubating homogenate on ice; centrifugally separating, centrifuging at low temperature and medium speed, taking precipitate, and discarding supernatant; core washing, re-suspending the precipitate in a washing buffer solution, uniformly mixing and filtering; performing core extraction, re-suspending the precipitate in a suspension buffer solution, and lightly and uniformly mixing; purifying and washing, carrying out density gradient centrifugation on the suspension, dyeing with trypan blue, taking a core layer, and washing for subsequent experiments. The nuclear extraction lysis buffer solution has two proportions of LB1 and LB2, different tissues are suitable for different proportions, and an enzyme inhibitor can be added as required. The method can be used for rapidly extracting various animal tissue cell nucleuses, is complete in shape, large in quantity and few in impurities, is suitable for different tissues, and can be applied to downstream experiments such as single cell nucleus sequencing.
Owner:HANGZHOU CHANGSHENG FRUIT GENE TECHNOLOGY CO LTD

A kit and method for extracting plant tissue nuclei

This invention discloses a kit and method for extracting cell nuclei from plant tissues, belonging to the field of biotechnology. The kit includes a nuclear lysis buffer and a nuclear washing buffer. The nuclear lysis buffer is prepared using nuclease-free water with sucrose, Tris-HCl (pH 7-8), NaCl, MgCl2, a surfactant, DTT, an RNase inhibitor, and spermidine. The nuclear washing buffer is prepared using nuclease-free water with sucrose, Tris-HCl (pH 7-8), NaCl, MgCl2, DTT, an RNase inhibitor, and spermidine. This kit is suitable for extracting cell nuclei from fresh yew stem tissue, yielding cell nuclei with intact nuclear membranes, stable gene expression, high purity, and few impurities, suitable for single-cell sequencing.
Owner:HANGZHOU LC BIOTECH

Human glioma organoid isolated culture kit and human glioma organoid culture method

The invention belongs to the technical field of regenerative medicine and biology, and particularly relates to a human glioma organoid isolated culture kit and a human glioma organoid culture method. The human glioma organoid isolated culture kit comprises a tissue activity protection solution, a P1 buffer solution, a human glioma organoid culture medium, an organoid cryopreservation solution and a red blood cell lysis buffer, and the culture method is carried out by adopting the human glioma organoid isolated culture kit. The human glioma organoid can be cultured by adopting the human glioma organoid isolated culture kit and the culture method, and the kit and the culture method have the advantages that the human glioma organoid grows quickly and can be cryopreserved and resuscitated.
Owner:SOUTH CHINA NORMAL UNIV

Cracking buffer solution, kit and extraction method for extracting free DNA (Deoxyribose Nucleic Acid) from peripheral blood

The invention relates to the technical field of nucleic acid detection, in particular to a lysis buffer solution, a kit and an extraction method for extracting free DNA of peripheral blood. The lysis buffer solution for extracting the free DNA of the peripheral blood comprises a first lysis solution and a second lysis solution, the first lysis solution takes guanidine hydrochloride as a main chaotropic agent and at least one of ammonium thiocyanate and guanidine isothiocyanate as an auxiliary chaotropic agent, and is compounded with a metal ion chelating agent, trehalose, dithiothreitol, digestive enzyme, sodium dodecyl sarcosinate, borate and ascorbic acid; the second lysis solution takes guanidine hydrochloride as a main chaotropic agent and urea as an auxiliary chaotropic agent, and is compounded with a metal ion chelating agent, trehalose, dithiothreitol, digestive enzyme, sodium dodecyl sarcosinate, borate, ascorbic acid and polyethylene glycol. The buffer solution of the lysate does not contain SDS (sodium dodecyl sulfate), so that the residue of SDS in an extracted product can be avoided, and the amount of extracted free DNA (deoxyribonucleic acid) is more.
Owner:SHANGHAI JINFUKANG PHARMACEUTICAL ENGINEERING TECHNOLOGY CO LTD

Process for the preparation of desipramine

This invention relates to the field of peptide preparation technology, specifically to a preparation process for terfenadine. The terfenadine preparation process includes the following steps: (1) using resin as a solid-phase carrier, Boc-Pip(Fmoc)-OH is loaded onto the resin to obtain Boc-Pip(Fmoc)-resin; (2) under the action of a condensing agent, Fmoc-D-Lys(Boc)-OH and Boc-D-Phe-D-Phe-D-Leu-OH are sequentially coupled onto the Boc-Pip(Fmoc)-resin to obtain terfenadine peptide resin; (3) the terfenadine peptide resin is lysed using a lysis buffer, then concentrated, precipitated, and dried to obtain crude terfenadine. This invention's preparation process can effectively improve product purity while ensuring large-scale industrial production, and the steps are simple with a short reaction cycle.
Owner:CHENGDU SINTANOVO BIOTECHNOLOGY CO LTD