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48 results about "Protease K" patented technology

IV type collagen detection kit and preparation method thereof

The invention discloses an IV-type collagen detection kit and a preparation method thereof, and relates to the technical field of in vitro diagnos.The kit comprises a reagent R1 and a reagent R2, and the volume ratio of the reagent R1 to the reagent R2 is 3: 1; through an elaborately designed reagent formula, especially the accurate proportion of all components in the reagent R1, such as ammonium chloride, sodium azide and Brij-35, the detection sensitivity and accuracy are improved, the stability and repeatability of the kit are also ensured, and particularly, the natural plant extract astragalus polysaccharide and the protease K inhibitor are introduced into the reagent R1 as key components, so that the detection sensitivity and accuracy are improved, and the detection sensitivity and repeatability are improved. According to the present invention, the unique preparation method and the activity promotion effect on the IV-type collagen hydrolase significantly enhance the biological activity of the kit, such that the detection result is reliable, and the two latex particles coated with the paired mouse monoclonal antibodies adopted in the reagent R2 have high specificity and affinity, such that the kit can be used for the detection of the IV-type collagen hydrolase. And the detection accuracy and sensitivity are further improved.
Owner:JIANGSU MAIYUAN BIOTECHNOLOGY 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

Method for detecting Tween 20 in protein preparation

The invention relates to a colorimetric detection method for Tween-20 in a protein preparation, which comprises the following steps: diluting the protein preparation with a diluent for enzymolysis, carrying out chromogenic reaction by using dichloromethane and an ammonium iron thiocyanate system, and measuring absorbance by using a visible spectrophotometer. The diluent is composed of other components except protein and Tween 20 in the protein preparation; an enzyme used in the enzymolysis is protease K; the concentration of the protease K is preferably 200 mu g / ml; the enzymolysis temperature is preferably 55-57 DEG C; the enzymolysis time is preferably 3.5 to 4 hours.
Owner:CHENGDU KANGHONG BIOTECH CO LTD

Nucleic acid extraction method and nucleic acid extraction kit

The invention relates to the technical field of nucleic acid extraction, in particular to a nucleic acid extraction method and a nucleic acid extraction kit. The nucleic acid extraction kit can remove protein in a sample and further release / purify nucleic acid. According to the nucleic acid extraction method, the protease K is added in the washing liquid of the extraction reagent, protein which is deeply combined can be treated, nucleic acid is further released, high-temperature dissociation is used in the nucleic acid elution process, the protease K is inactivated, and the influence on subsequent amplification is avoided.
Owner:AUTOBIO DIAGNOSTICS CO LTD

High-sensitivity peripheral blood free DNA magnetic bead capture system, methylation transformation method kit and application

The invention relates to the technical field of molecular diagnosis, in particular to a high-sensitivity peripheral blood free DNA magnetic bead capture system, a methylation transformation method kit and application. The kit is composed of a lysis solution, a protease K diluent, a composite conversion reagent, a washing solution, a scavenger, an eluent and combined magnetic beads. According to the high-sensitivity peripheral blood free DNA magnetic bead capture system and the methylation transformation method kit, the detection limit of early-stage lung cancer ctDNA reaches 0.005% AF, the I-stage cancer ctDNA detection rate is increased to 93% from the industry average 40%, the sample size is reduced to 4 mL from the conventional 10 mL, the batch-to-batch difference of the kit is small, and the kit has very high application value clinically.
Owner:SUZHOU CHIEN SHIUNG INST OF TECH

Cosmetic composition containing triple polishing enzyme and preparation method thereof

The invention relates to the field of cosmetic compositions, in particular to a cosmetic composition containing triple polishing enzyme and a preparation method of the cosmetic composition. The triple polishing enzyme comprises protease, protease K and superoxide dismutase, the weight part ratio of the protease to the protease K to the superoxide dismutase is 1: (0.5-2): (0.5-2), the total mass percent of the triple polishing enzyme in the composition is 0.0002%-0.001%, and the invention further discloses a preparation method of the cosmetic composition. A triple polishing enzyme synergistic system is formed through scientific compounding, the integrated skin care effect under low concentration is achieved, and the defects of a traditional chemical stripping agent and the shortages of a single enzyme preparation are overcome; meanwhile, by optimizing the proportion of formula components, the composition forms a liquid crystal structure, so that long-acting dual moisturizing of internal water locking and external water sealing is realized, triple polishing enzyme can be wrapped and protected, and the problems that the composition is easy to oxidize and inactivate and low in transdermal efficiency are solved.
Owner:EIIO COSMETICS (ZHEJIANG) CO LTD +2

Method for efficiently extracting genome DNA (Deoxyribose Nucleic Acid) of Snaketail

The invention discloses a method for efficiently extracting genome DNA (Deoxyribose Nucleic Acid) of ophiurocephalus serratus. The method comprises the following steps: mixing a sample with a lysis solution and metal balls, crushing the mixture at-20 to-10 DEG C for 250-350s at 40-60Hz, and placing the crushed sample at-20 to 4 DEG C; cracking the sample at the temperature of 50-60 DEG C for 4-6 minutes; centrifuging at 2-4 DEG C, taking a supernatant, adding an extraction agent, and uniformly mixing; centrifuging at 2-4 DEG C, taking an upper-layer solution, adding an extracting agent, and uniformly mixing; centrifuging at 2-4 DEG C, taking an upper-layer solution, adding isopropanol, uniformly mixing, and standing at-20--15 DEG C for 20-30 minutes; centrifuging at 2-4 DEG C, discarding the supernatant, washing the precipitate with an ethanol solution, and dissolving with an eluent; the lysis solution contains CTAB (Cetyltrimethyl Ammonium Bromide), protease K and beta-mercaptoethanol. According to the invention, the problem of high-mineralization echinodermis sample treatment is solved, and a high-quality nucleic acid template and a new technology are provided for research on ocean invertebrate genetics.
Owner:SOUTH CHINA SEA INST OF OCEANOLOGY CHINESE ACAD OF SCI

Narrow-spectrum antibacterial peptide XMK-8 and application thereof

The invention belongs to the technical field of biology, and relates to a narrow-spectrum antibacterial peptide XMK-8 and application thereof. The amino acid sequence of the narrow-spectrum antibacterial peptide XMK-8 is as shown in SEQ ID NO: 1, and the narrow-spectrum antibacterial peptide XMK-8 has no bacteriostatic activity on escherichia coli, salmonella, staphylococcus aureus and aeromonas hydrophila and only has bacteriostatic activity on pasteurella multocida and haemophilus parasuis. The antibacterial activity of the narrow-spectrum antibacterial peptide XMK-8 can still be detected after treatment with salt ions (sodium ions and potassium ions, 50-200 mmol / L), acid-base (pH 4-10) and protease K (20-100 [mu] g / mL) at the temperature of 0-100 DEG C, and the narrow-spectrum antibacterial peptide XMK-8 has low hemolytic activity on red blood cells of mice. The antibacterial peptide XMK-8 disclosed by the invention is expected to become a novel antibiotic substitute and has a good application prospect in prevention, treatment, diagnosis and identification of pasteurella multocida, haemophilus parasuis and infection thereof.
Owner:HENAN INST OF SCI & TECH

Method for detecting methylation level of heterozygous heterologous DNA in saccharomyces cerevisiae

The invention discloses a method for detecting methylation level of heterozygous heterologous DNA (deoxyribonucleic acid) in saccharomyces cerevisiae. Comprising the following steps: preparing a saccharomyces cerevisiae genome, carrying out enzymolysis on the saccharomyces cerevisiae genome, and detecting the methylation level of heterozygous heterologous DNA in saccharomyces cerevisiae by using an ultra-high performance liquid chromatography-triple quadrupole mass spectrometer. According to the method, saccharomyces cerevisiae cell walls can be better removed, the use time of protease K is adjusted, protein combined with DNA molecules is better removed, saccharomyces cerevisiae genomes are extracted with high quality, and impurities are greatly reduced; the PMSF protease inhibitor is added in the saccharomyces cerevisiae genome extraction process, so that the activity of protease K can be effectively inhibited, the residual protease K is prevented from degrading nuclease and other key enzymes in the subsequent DNA enzymolysis process, and the DNA enzymolysis efficiency is improved. The method can be directly applied to DNA methylation detection of saccharomyces cerevisiae.
Owner:TIANJIN UNIV

Method for extracting cattle genome from vegetable tanned cattle leather cultural relics

The invention relates to the field of DNA extraction of leather cultural relics, and discloses a method for extracting cattle genomes from vegetable tanned cattle leather cultural relics. The method comprises the following steps: after cleaning the vegetable tanned cow leather cultural relics, dehydrating, freeze-drying and grinding and crushing with liquid nitrogen; metal ions are chelated through an EDTA-Tris detanning reagent to destroy a tannin structure, so that the wrapping on DNA (Deoxyribose Nucleic Acid) is reduced; then, lysate and protease K are used for lysis and digestion, cell membranes are damaged, protein is degraded, and DNA is released; adding absolute ethyl alcohol to precipitate and separate out DNA; and finally, purifying, recovering and purifying the DNA through an adsorption column. The genome DNA extracted by the method is large in fragment, high in purity and stable and reliable in quality.
Owner:ZHEJIANG SCI-TECH UNIV

Polypeptides having angiotensin converting enzyme inhibitory activity, methods of making and use thereof

The application provides a polypeptide with angiotensin converting enzyme inhibitory activity and a preparation method and application thereof, and relates to the technical field of active peptide preparation. The abalone is pulverized, a complex enzyme is added for primary enzymolysis, and then a specific protease mixture containing prolyl endopeptidase, chymotrypsin and protease K is added for two-step enzymolysis, so that the obtained polypeptide solution has high ACE inhibitory activity. The obtained polypeptide is analyzed and screened to obtain polypeptides with amino acid sequences as shown in SEQ ID NO. 1-5. The polypeptide has high ACE inhibitory activity, and the IC 50 0.05mg / mL-0.40mg / mL, and therefore can play a blood pressure lowering effect as an angiotensin converting enzyme inhibitor.
Owner:JIANGNAN UNIV

Preparation method of nucleic acid detection sample

The invention provides a preparation method of a nucleic acid detection sample, which comprises the following steps: adding a pH response type hydrogel into an inactivated virus preservation solution, mixing, applying the inactivated virus preservation solution to virus sample collection to obtain a sample solution, applying the inactivated virus preservation solution to a nucleic acid detection sample, and detecting the nucleic acid detection sample according to the sample solution, the pH response range of the pH response type hydrogel is 5-6, and the pH value of the inactivated virus preservation solution is 7.5-8. Adding protease K and magnetic beads into the sample solution, uniformly mixing, performing enzymolysis, adjusting the pH value to 5-6, incubating for 10-30 minutes, centrifuging a container filled with the sample solution, fixing the magnetic beads at the bottom of the container through a magnet, removing supernate, washing the magnetic beads at least once, and drying the washed magnetic beads to obtain a sample solution; and eluting the washed magnetic beads to obtain the nucleic acid detection sample. According to the preparation method, a nucleic acid detection sample with higher nucleic acid content can be obtained, so that the accuracy of nucleic acid detection is improved.
Owner:NANJING FANGHUA GENE TECH CO LTD

Extraction reagent, kit and extraction method for free DNA (Deoxyribose Nucleic Acid) of plasma

The invention discloses a plasma free DNA extraction reagent, a kit and an extraction method, the extraction reagent comprises protease K, a lysis solution, a binding solution, a washing solution 1, a washing solution 2, a washing solution 3 and an eluent, the extraction reagents contain more common and less-dosage common reagents such as Tris-HCl (pH 8.0), EDTA.2Na. 2H2O, NaCl, glycerol, SDS, isopropanol, ethanol, Tween-20 and the like. The method has the advantages that the problems that the reagent is easy to separate out and needs to be heated and redissolved for use during storage can be solved without traditional high-concentration guanidine salts such as guanidine hydrochloride and guanidinium isothiocyanate, the stability and the operation convenience of the extraction reagent are increased, a better extraction effect is achieved, and the eluent can effectively protect the extracted cfDNA from being degraded.
Owner:ONKAI LIFE SCIENCES (SUZHOU) CO LTD

Detection method for determining form of trace selenium element in geological high background region vegetables

The invention belongs to the technical field of food detection, and particularly relates to a detection method for determining the form of trace selenium in vegetables in a geological high background area. The method comprises the following steps: cleaning a vegetable sample, freeze-drying, grinding and crushing, adding a specific compound enzyme system (protease XIV, protease K and cellulase) and a Tris-HCl buffer solution, performing water bath ultrasonic treatment, performing constant-temperature oscillation for enzymolysis, performing low-temperature centrifugation on an extracting solution for multiple times, and filtering and purifying the extracting solution by a microporous filter membrane, the method comprises the following steps: detecting the selenium form of a sample by adopting a high performance liquid chromatograph and inductively coupled plasma mass spectrometer serial device, and quantitatively analyzing tetravalent selenium (Se4 +), hexavalent selenium (Se6 +), selenomethionine (SeMet), methylselenocysteine (SeMeCys) and selenocystine (SeCys2) in the sample by adopting a normalization method. The detection method disclosed by the invention is strong in anti-interference capability and high in sensitivity, and original form information can be completely reserved.
Owner:广东省地质调查研究院 +1

Method for extracting plant RNA (Ribonucleic Acid)

The invention discloses a plant RNA extraction method, and belongs to the technical field of RNA extraction. The extraction method of the plant RNA comprises the following steps: mixing plant tissue powder, a CTAB lysate, a sulfydryl reducing agent and protease K, and then cracking at 60-65 DEG C to obtain a lysate; centrifuging the lysate, taking a supernatant, mixing the supernatant with a first solvent, oscillating, and washing the supernatant to obtain a precipitate; the precipitate and nuclease-free water are mixed and dissolved, and then DNase I and Buffer RDD are added for incubation; continuously adding sodium acetate, an RNA extraction phenol reagent and a second solvent, oscillating, carrying out ice bath, and centrifuging to obtain supernate; carrying out isopropanol precipitation on the supernate to obtain plant RNA (Ribonucleic Acid); the first solvent or the second solvent is a mixture of chloroform and isoamyl alcohol. According to the extraction method provided by the invention, the purpose of completely digesting the genome DNA is achieved on the premise of not influencing the quality of extracted RNA.
Owner:WUHAN FRASERGEN CO LTD

Immunofluorescence method and kit for detecting zebra fish heart DNA methylation level

The invention relates to an immunofluorescence method and kit for detecting zebra fish heart DNA methylation level, and belongs to the technical field of biological detection. The immunofluorescence method provided by the invention comprises the following steps: exposing a zebrafish embryo to a substance to be detected, dissecting a heart sample of the zebrafish embryo and carrying out fixation, membrane permeation, protease K digestion, hydrochloric acid DNA denaturation and neutralization treatment, and then carrying out sealing, incubation of a 5-methylcytosine primary antibody and a fluorescently labeled secondary antibody, cell nucleus staining and fluorescence detection. And finally, analyzing and evaluating the DNA methylation level through fluorescence imaging. According to the method disclosed by the invention, through a combined step of protease K and hydrochloric acid denaturation treatment, the binding efficiency to the antibody is remarkably improved, so that high-sensitivity and low-background detection on the methylation level of the whole DNA in a zebra fish micro heart sample is realized; the detection kit provided by the invention can be widely applied to epigenetics evaluation of toxicity of environmental pollutants.
Owner:SUZHOU UNIV

Polypeptide with angiotensin converting enzyme inhibition and preparation method and application thereof

The invention provides a polypeptide with angiotensin converting enzyme inhibition as well as a preparation method and application thereof, and relates to the technical field of active peptide preparation. According to the method, the abalone is crushed, the compound enzyme is added for primary enzymolysis, then the specific protease mixture containing prolyl endopeptidase, chymotrypsin and protease K is added for secondary enzymolysis, and the obtained polypeptide liquid has high ACE (angiotensin converting enzyme) inhibition. The obtained polypeptide is analyzed and screened to obtain the polypeptide with the amino acid sequence as shown in SEQ ID NO.1-5. The polypeptide disclosed by the invention has high ACE (angiotensin converting enzyme) inhibition, and IC50 (50% of IC50) of the polypeptide to ACE is 0.05 mg / mL-0. 40mg / mL, so that the polypeptide can be used as an angiotensin converting enzyme inhibitor to play a role in reducing blood pressure.
Owner:JIANGNAN UNIV

DNA / RNA extraction kit based on magnetic bead method without enzyme digestion

The application discloses a DNA / RNA reagent kit based on a magnetic bead method and free of enzyme digestion, and belongs to the technical field of nucleic acid extraction and purification. The reagent kit comprises a lysis solution AL, protease K, a specific binding buffer CLB, amino magnetic bead suspension, silicon hydroxyl magnetic bead suspension and other components. The application solves the problems of nucleic acid degradation, incomplete removal of cross contamination and complicated process caused by enzyme digestion in the prior art, is suitable for automatic high-throughput processing, and has a wide application prospect in scientific research and clinical application.
Owner:GUANGZHOU BAYBIO BIO-TECH CO LTD

Lysate for candida albicans, kit and gene detection method

The invention belongs to the technical field of biomedical detection, and particularly relates to a lysis solution for candida albicans, a kit and a gene detection method. The cleavage efficiency and background interference are balanced by regulating and controlling the concentration of saponin, nucleic acid can be wrapped by too many saponin micelles, and cleavage is incomplete if too few saponin micelles exist; citric acid buffers to adjust the acidic environment, although RNA degradation may be promoted, nuclease is cleared through high-salt rapid cracking and protease K, and negative effects are counteracted; the pre-activated protease K is synchronous with membrane rupture, so that the enzyme activity peak is matched with the exposure time of the target protein, the degradation efficiency is improved, and a cracking technology which can simultaneously realize high-yield and high-function nucleic acid output and is adaptive to a rapid detection process is developed.
Owner:LANZHOU BAIYUAN GENE TECH

Nucleic acid hybrid capture immunofluorescence detection kit as well as use method and application thereof

The invention belongs to the technical field of molecular detection, and discloses a nucleic acid hybridization capture immunofluorescence detection kit as well as a use method and application thereof. In the kit provided by the invention, guanidine hydrochloride, guanidine isothiocyanate, triton X-100, NaCl, protease K and lauryl sodium sulfate are compounded according to a specific concentration to obtain the nucleic acid release liquid, so that pathogen nucleic acid in a clinical sample can be fully dissociated, and the nucleic acid release liquid can cooperate with a probe aiming at a specific target pathogen; the solution system can be used for optimizing the recognition and combination process of DNA and RNA, the specific recognition and combination capacity between the probe and specific pathogen nucleotide is improved, and under the condition that a common nucleic acid stabilizer, namely Tween, is not contained, the solution system can further strengthen differential structural epitopes existing in a DNA / RNA complex compared with DNA and RNA, so that the specificity of the DNA / RNA complex is improved, and the specificity of the DNA / RNA complex is improved. The kit can be better recognized and combined by an anti-DNA / RNA complex antibody, and has excellent detection specificity, sensitivity and precision.
Owner:ANBIO XIAMEN BIOTECHNOLOGY CO LTD

Marine organism muscle tissue DNA extraction kit and application thereof

PendingCN122629172AMuscle tissueLysis
The application discloses a marine organism muscle tissue DNA extraction kit and application thereof, and belongs to the technical field of biological detection. The extraction kit comprises a lysis solution, a first washing solution and a second washing solution. The lysis solution is composed of Triton x-100 45 mM, Tris-HCl 10 mM, EDTA-Na 80 mM, NaCl 80 mM and Brij-35 3 mM, and has a pH of 8.3. The first washing solution is composed of 70% ethanol solution containing 200 mM Tris-HCl. The second washing solution is composed of 80% ethanol solution. The marine organisms include oceanic fishes, Antarctic fishes, shellfishes, crabs and shrimps. The tissue is taken, cut, mixed with the lysis solution and protease K, and uniformly mixed in a water bath. The application is based on the phenol extraction method, and various mild reagents are mixed to realize rapid and accurate DNA extraction, so that the purity, quality and integrity of the DNA are ensured.
Owner:SHANGHAI OCEAN UNIV

Extraction method of trace phyllanthus emblica plant tissue total RNA

The invention discloses an extraction method of trace phyllanthus emblica plant tissue total RNA, and belongs to the technical field of plant molecular biology. The extraction method provided by the invention is established by preparing an extraction reagent from sodium borate decahydrate, EGTA, SDS, DTT, PVP and NP-40 and combining experimental methods such as protease K enzymolysis, lithium chloride precipitation, potassium acetate purification and absolute ethyl alcohol precipitation. Experimental results show that the extraction method provided by the invention can improve the purity and stability of RNA, and has the characteristics of low cost, low pollution, high flexibility and high universality. The invention provides a reliable scheme for extracting the total RNA of different tissue plant samples of the phyllanthus emblica, and lays an early-stage method foundation for molecular biology research, transcriptome analysis, gene expression research and the like of the phyllanthus emblica in the future.
Owner:YUNNAN ACAD OF AGRI SCI +1

Automatic body fluid sample pathogen and host genome nucleic acid synchronous extraction method

The invention discloses an automatic body fluid sample pathogen and host genome nucleic acid synchronous extraction method. The method comprises the following steps: S1, mixing a large-volume body fluid sample with a lysis buffer solution Buffer LA containing chaotropic salt and protease K, and carrying out lysis to release nucleic acid; s2, amino-modified magnetic beads are added, nucleic acid is captured through electrostatic interaction, after room-temperature combination, magnetic separation is conducted, supernatant is abandoned, and the amino-modified magnetic beads are reserved; s3, eluting nucleic acid on the amino-modified magnetic beads, and performing magnetic separation to retain eluent; s4, adding a protective agent Buffer GTN, isopropanol and ion exchange magnetic beads into the eluent, uniformly mixing and combining, performing magnetic separation, discarding supernatant, and retaining the ion exchange magnetic beads; s5, washing the ion exchange magnetic beads, drying, and eluting with Buffer EB1 to obtain whole nucleic acid; according to the method, large-volume, high-efficiency and high-purity nucleic acid synchronous extraction is realized through a double-magnetic-bead staged capturing and cracking-protective agent combination technology.
Owner:ZHONGSHAN HOSPITAL FUDAN UNIV

Kit for extracting specific short fragment cfDNA in urine and application thereof

The invention discloses a kit capable of being used for extracting specific short fragment cfDNA in urine and application of the kit. The kit comprises protease K, a lysis solution, a cleaning solution, an eluent and magnetic beads, wherein the magnetic beads comprise silicon hydroxyl magnetic microspheres and magnetic microspheres modified by streptavidin. According to the technical scheme, through matching of composition control of the magnetic beads and optimization of selection of the components, when the obtained kit is used for extracting free DNA in urine, the extraction efficiency of specific short fragments is greatly improved, and the detection accuracy is further improved.
Owner:BEIJING BOE TECH DEV CO LTD +1

Ultrahigh sensitivity RNA in-situ detection method

The invention discloses an ultrahigh sensitivity RNA in-situ detection method. The method comprises the following steps: S1, obtaining a tissue slice through a frozen section; s2, fixing is conducted for 15 min through 4% PFA; s3, 2x SSC rinsing is carried out twice, and each rinsing is carried out for 5 min; s4, rinsing with 70% alcohol; s5, soaking in 70% alcohol for 2 hours; s6, carrying out # imgabs0 # treatment for 15 minutes; s7, PBST rinsing is carried out once; s8, protease K treatment is performed for 10 min; s9, rinsing twice with 4% BSA (Bovine Serum Albumin); s10, 2x SSC rinsing is carried out twice; s11, sealing is conducted for 30 min through sealing liquid; s12, performing probe hybridization for 2 hours; s13, cleaning with a cleaning solution twice, 0.5 h each time; s14, incubation is carried out for 0.5 h through stepdavidin-hrp (stem davidin-human recombinant protein); s15, 4% BSA cleaning is carried out five times, and each time is 5 min; s16, carrying out a TSA reaction for 15 min; s17, sheet sealing is carried out; and S18, observing and recording the signal through a microscope. The probe is divided into the primary probe and the secondary probe, the primary probe and the secondary probe are mixed to form the complete probe, a plurality of probes are designed for each RNA molecule, signal amplification is carried out, and signal amplification is carried out through combination of streptavidin coupled with HRP and biotin of the secondary probe and through a Tyramide signal amplification technology, so that signal detection with ultrahigh sensitivity can be achieved.
Owner:HANGZHOU RUIU BIOTECHNOLOGY CO LTD

Cat polycystic kidney gene mutation detection kit based on CRISPR-Cas12a

The invention relates to the technical field of biological detection, in particular to a cat polycystic kidney gene mutation detection kit based on CRI SPR-Cas12a. The kit comprises a freeze-dried CRI SPR reaction ball which is composed of a Cas12a enzyme, a targeted PKD1 gene c.10063Cgt, a CRI SPR gene c.10063Cgt, a CRI SPR gene c.10063Cgt and a CRI SPR gene c.10063Cgt. The kit is formed by premixing and freeze-drying gRNA of a T mutation site and a fluorescence labeling-quenching group reporter probe. The normal-temperature lysis solution is prepared from protease K, a TritonX-100 surfactant and sodium chloride; the chromogenic test strip is formed by arranging two detection lines, namely a T1 line and a T2 line, on a nitrocellulose membrane; and a quality control line C. The CRI SPR reaction ball is freeze-dried to integrate Cas12a enzyme, PAM simulates gRNA and double reporter probes, so that the normal-temperature long-acting stability of key components is realized; a normal-temperature lysis solution is fused with thermal stable protease K and a nuclease inhibition technology, sample DNA is directly released in a short time, degradation is blocked, and the step of centrifugal purification is eliminated.
Owner:SUZHOU JIDITAI BIOTECHNOLOGY CO LTD

Dyeing reagent and dyeing method applied to acid-fast dyeing

The invention belongs to the technical field of medical detection, and discloses a staining reagent applied to acid-fast staining and a staining method. The dyeing reagent comprises a primary dyeing agent, a decolorizing agent, a counterdyeing agent and an accelerating agent, wherein the accelerating agent is prepared from the following components: lauryl sodium sulfate, triton X-100, hexadecyl trimethyl ammonium bromide, a Tris-HCl buffer solution, ethylenediamine tetraacetic acid, sodium chloride, potassium chloride, protease K and beta-mercaptoethanol. According to the dyeing reagent, the dyeing accelerant is introduced into a traditional acid-fast dyeing system, the component capable of specifically damaging the lipid structure of the cell wall of the Mycobacterium is utilized, the hindering effect of the lipid structure on the dye in the dyeing process is reduced, the binding efficiency of the dye and thalli is remarkably improved, and therefore the positive detection rate of acid-fast dyeing is greatly increased.
Owner:广州市胸科医院

Preparation method of fluorescence in situ hybridization kit based on hybridization chain reaction (HCR)

The invention belongs to the technical field of molecular biology, and discloses a preparation method of a fluorescence in situ hybridization kit based on a hybridization chain reaction (HCR), the core components of the kit comprise a hybridization probe, an amplification probe, a washing liquid, a confining liquid, protease K, HRP-mouse anti-digoxin, and a TSA developing liquid 520 nm. According to the FISH detection system combined with the HCR signal amplification technology, when a hybridization probe and an amplification probe are simultaneously combined to a position adjacent to a target sequence, a small section of sequence at the top end can trigger a branched HCR reaction to form a macromolecular nucleic acid aggregate. Signal amplification of HCR is achieved through cascade hybridization reaction, detection sensitivity and specificity can be remarkably improved, and meanwhile the problem of background noise caused by a traditional enzymatic method is avoided. The kit disclosed by the invention is wide in sample application range, and can be used for positioning analysis of target RNA of samples such as paraformaldehyde or formalin-fixed animal and plant tissues, paraffin or frozen sections, cells and the like, and high-sensitivity detection of the single cell level of non-coding RNA.
Owner:HANGZHOU BORRICK BIOTECHNOLOGY CO LTD

Targeted protein degradation robot based on DNA origami and construction method and application thereof

The invention discloses a targeted protein degradation robot based on DNA origami as well as a construction method and application of the targeted protein degradation robot, and relates to the technical field of biological medicines. The targeted protein degradation robot comprises a DNA origami structure and a recognition module, the construction method of the DNA origami structure comprises the following steps: mixing M13 ssDNA with 60 stapled chains, carrying out annealing reaction to obtain DNA six-screw column origami, and carrying out incubation reaction on the DNA six-screw column origami and a P chain coupled with protease K to obtain origami coupled with inner layer protease K; performing incubation reaction on the origami coupled with the inner layer protease K, a T + S + A-ABD compound and an L + A-ABD compound to obtain an origami compound; and carrying out incubation reaction on the origami compound and human serum albumin (HSA) to obtain the DNA origami structure. The robot can achieve non-endogenous-dependence in-situ precise degradation of target protein, and a new strategy is provided for achieving treatment of diseases such as tumors through interference of the protein level.
Owner:EAST CHINA UNIV OF SCI & TECH

Preparation method and application of rice callus-derived pdrn

The application discloses a preparation method and application of PDRN (plant-derived RNA) from rice callus. The preparation method comprises the following steps: inducing and suspending culturing of the rice callus; homogenizing the rice callus suspension cells obtained in step 1; sequentially using cellulase, pectinase and protease K for enzymolysis; nucleic acid purification and precise ultrasonic shearing are performed in sequence, so that PDRN with a molecular weight of 25-500 bp is obtained. Through optimization of the rice callus culture condition and combination of the joint lysis process of "mechanical grinding-composite enzymolysis-protease treatment", efficient and high-purity preparation of the plant source PDRN is realized, the extraction rate can be higher than 0.5 mg / g of fresh weight of the callus, and the purity A260 / A280 is greater than or equal to 1.8. The obtained PDRN has significant effects in terms of anti-wrinkle, soothing, moisturizing, anti-inflammatory and anti-aging.
Owner:GUANGDONG COOPERATE BIOTECHNOLOGY CO LTD