65 results about "RNA extraction" patented technology

The invention discloses a key gene identification method related to tobacco nitrogen response, which comprises the following steps: S1, setting four nitrogen fertilizer gradient treatments on the basis of same phosphorus and potassium fertilization by adopting a field experiment of a completely random block; s2, randomly taking the 6th to 8th leaves of the five plants in each area, and dividing a sample into two parts: quickly freezing one part with liquid nitrogen, and storing at-80 DEG C for RNA (Ribonucleic Acid) extraction and transcriptome sequencing; one part is used for measuring the nitrogen content, and after baking and drying treatment, a KjeltecTM8100 automatic nitrogen analyzer is used for measuring; s3, nitrogen content determination: determining the nitrogen content in the treated sample by using an automatic nitrogen analyzer KjeltecTM8100; according to the method, high-throughput transcriptome sequencing and weighted gene co-expression network analysis (WGCNA) are combined, so that not only can gene expression maps of flue-cured tobacco leaves treated by different nitrogen fertilizers be comprehensively captured, but also gene modules with similar expression modes can be mined from a global perspective.

The invention discloses a composition for detecting benign and malignant thyroid nodules and a detection method thereof, and relates to the technical field of medical detection, the composition comprises the following components: a DNA / RNA extraction reagent and a fluorescent quantitative PCR reagent; by combining the DNA / RNA extraction reagent and the fluorescent quantitative PCR reagent, nucleic acid can be efficiently and accurately extracted from the thyroid nodule fine needle puncture sample, specific gene mutation and gene fusion are detected through the fluorescent quantitative PCR technology, the accuracy and sensitivity of benign and malignant judgment of the thyroid nodule are remarkably improved, and the method is suitable for clinical application. And a doctor can formulate a treatment scheme earlier and more accurately, and the prognosis of a patient is improved.

This RNA extraction method comprises: (1) adding an alkali metal salt and a cell wall-degrading enzyme to a sample to prepare a reaction solution containing the alkali metal salt and the cell wall-degrading enzyme, the final concentration of the alkali metal salt being 50 mM to 150 mM; (2-1) incubating the reaction solution to degrade the cell walls of microorganisms contained in the reaction solution; and (3) extracting RNA from the reaction solution after the incubation. This reagent set for RNA extraction contains: an alkali metal salt solution in which the concentration of the alkali metal salt is 50 mM or more; and a cell wall degrading enzyme.

The invention provides a DNA library screening method and application of an intracellular mRNA sensor switch. The DNA library screening method comprises the following steps: designing a DNA library containing a switch sequence of a degenerate basic group; preparing a circular expression vector library; preparing a minimum linear expression vector library; performing seamless cloning; carrying out RCA rolling circle amplification; carrying out telomerase digestion; carrying out cell transfection; extracting RNA (Ribonucleic Acid) and carrying out reverse transcription; carrying out library building of next-generation sequencing by taking the cDNA as a template; carrying out next-generation sequencing; analyzing data and comparing to obtain the DNA library of the mRNA sensor switch. According to the DNA library screening method provided by the invention, more sensor switch sequences can be provided for a single site on mRNA in cells, and more selection possibilities are further provided for subsequent targeted therapy.

The invention relates to the field of gene detection, and particularly discloses a gene detection device and method applying Beidou satellite positioning, and the device comprises a sample collection and preprocessing module which is used for obtaining a body fluid or tissue biological sample of a user, and carrying out cell lysis, DNA / RNA extraction and purification operation on the sample; the gene sequencing analysis module is connected with the sample collecting and preprocessing module and is used for performing gene sequencing on the purified nucleic acid sample and identifying gene variation information related to tumors; the high-precision positioning capability of the Beidou system is utilized to obtain the environmental parameters of the geographic position of the user in real time, then the genetic variation data and the regional environmental carcinogenic factors are subjected to weighted fusion analysis through the risk assessment algorithm, and finally a highly personalized detection report is generated. In this way, the detection result can truly reflect the specific influence of the external environment on the individual health, and the accuracy and practicability of the report are remarkably improved.

The invention discloses a group of hydroxyl magnetic bead extraction and capture probes for GBSRNA, a kit and an extraction method, and relates to the technical field of nucleic acid extraction. The capture probe provided by the invention comprises five nucleotide sequences, the five nucleotide sequences are loaded on a hydroxyl magnetic bead and are used for preparing the kit for extracting the GBSRNA, the extraction efficiency of the RNA is greatly improved by rapidly splitting bacteria and releasing the RNA and specifically capturing the GBSRNA by utilizing the probe on the magnetic bead, and finally, DNA impurities are removed by utilizing DNase, so that the high-purity RNA is obtained. The technical problem that the extraction efficiency of RNA is relatively low due to relatively high content of impurities in an extract caused by inaccurate extraction is solved.

The application belongs to the technical field of RNA extraction devices, and particularly relates to an ep tube rack for RNA extraction, which comprises an ep tube rack body, a controller, a placing hole, a sucking assembly and a shaking assembly. A plurality of rows of placing holes with different specifications are formed in the ep tube rack body. The sucking assembly comprises a clamping ring, a fixing frame, a fixing column and a stabilizing frame. The fixing frame is fixed to one side of the front end of the ep tube rack body. The fixing columns are rotatably connected to the adjacent surfaces on the upper ends of the two fixing frames, and the fixing columns are damping shafts. The clamping ring is fixed between the two fixing columns. The stabilizing frame is fixed to the middle of the front end of the ep tube rack body. The application solves the problem that if the experimenter shakes hands, the upper layer of colorless water phase is easily transferred to a new ep tube after centrifugation, which can cause experimental failure.

The present application relates to the technical field of RNA extraction, and particularly relates to a pretreatment reagent, a method and a kit for RNA purification of whole blood samples. By introducing hydrophobic DESs as the core pretreatment component, the DESs are mixed with the whole blood sample in advance, the DESs form an independent phase layer after centrifugation by using the unique physicochemical properties, and the liposoluble drug components, inflammatory proteins and pigments are selectively extracted to the hydrophobic DESs phase and the RNA is preliminarily purified; the upper water phase of the pretreatment and purification is taken and added into a lysis solution containing a chaotropic salt to fully release the nucleic acid, and DNase I is introduced for digestion. The present application combines the pretreatment of hydrophobic DESs and the digestion of DNase I, and is used in combination with PVP (targeting adsorption of residual pigments and phenolic compounds), reduces the impurities of the lysis system through the front-end pretreatment, creates a low-interference and high-efficiency purification environment for the specific binding of RNA by the magnetic beads, and forms a synergistic removal network for the interference of complex samples.

The present invention relates to a kit for quantitatively examining mRNA of hair loss–related genes from hair, preferably from hair follicles, and a method for diagnosing or predicting causes of hair loss using same, and a method for alleviating, preventing, or treating hair loss. According to the present invention, testing from RNA extraction of hair loss–related genes to quantitative PCR can be performed using a small amount of hair, thereby enabling simple and rapid identification of the cause of hair loss and allowing for personalized hair health management. In addition, since no separate laboratory is required for on-site testing and anyone can easily perform testing and result acquisition through simple training, a user can easily perform the testing even in small clinics or facilities without clinical laboratories.

The application provides a DNA library screening method of intracellular mRNA sensor switch and application. The DNA library screening method comprises the following steps: designing a DNA library containing a switch sequence with a complementary base; preparing a circular expression vector library; preparing a minimized linear expression vector library; seamless cloning; RCA rolling circle amplification; telomerase enzyme cutting; cell transfection; RNA extraction and reverse transcription; library construction by performing second-generation sequencing with the cDNA as a template; second-generation sequencing; data analysis and comparison, and obtaining the DNA library of the mRNA sensor switch. The DNA library screening method provided by the application can provide more sensor switch sequences for a single site on intracellular mRNA, and further provides more selection possibilities for subsequent targeted treatment.

The invention belongs to the technical field of plant virus detection, and particularly relates to an RT-RPA-LFD method capable of rapidly and visually detecting three pathogens of sugarcane mosaic disease at the same time and application of the RT-RPA-LFD method. The invention relates to an RT-RPA-LFD primer group capable of simultaneously, rapidly and visually detecting three pathogens of sugarcane mosaic disease. The RT-RPA-LFD primer group comprises a primer group I, a primer group II and a primer group III, wherein the primer group I comprises an SrMV primer pair and an SrMV probe; the primer group II comprises an SCSMV primer pair and an SCSMV probe; and the primer group III comprises an SCMV primer pair and an SCMV probe. According to the multiple RT-RPA-LFD detection method disclosed by the invention, complicated RNA extraction, cDNA reverse transcription, gel electrophoresis and the like are not needed in the whole process, and multiple detection results can be observed by naked eyes within 5 minutes through a lateral flow test strip only by incubating at the constant temperature of 37 DEG C for 10 minutes. The detection system has the characteristics of rapidness, high efficiency, high sensitivity, strong specificity and no dependence on laboratory equipment, is suitable for on-site large-scale detection of field samples and sugarcane introduction materials, and greatly improves the detection efficiency.

The invention discloses a biological sample whole RNA normal-temperature long-term preservative as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The preservative comprises the following components: a tissue fixing agent, a tissue penetrating agent, a reactive biological sample encapsulating agent, a silicification blocking agent, a pH regulator, an ion concentration regulator and water. The preservative can be applied to bacteria, such as escherichia coli and staphylococcus aureus; a cell, such as an HEK293 cell, an ID8 cell; tissues such as mouse liver tissues and human tumor tissues; organs such as kidneys and hearts of small animals; the whole RNA of a complete organism, such as earthworms, fruit flies and other biological samples with different sizes can be stored at normal temperature for a long time, so that the RNA extraction efficiency and quality can be ensured, the method is particularly suitable for normal-temperature transfer and long-term storage of field collected samples and clinical submitted samples, and the RNA extraction efficiency and quality in subsequent molecular experiments can be ensured; the method is of great significance to molecular biology research and clinical diagnosis.

An RNA extraction kit comprises a kit body, a kit cover, a fixed seat, a movable plate, a pressing plate, a rotating sleeve, a shifting rod and a movable lock plate, the kit body is connected with the kit cover through a movable shaft rod, the movable shaft rod is sequentially connected with the kit body and the kit cover, the kit cover rotates on the movable shaft rod, the kit body is provided with a lock seat, the kit cover is provided with a spring bolt, the spring bolt is matched with the lock seat, and the spring bolt is connected with the lock seat. A fixed seat is connected inside the kit, the fixed seat is provided with a plurality of fixed grooves, the fixed grooves are matched with test tubes, the fixed grooves are connected with the test tubes, the test tubes are connected with test tube covers, a fixed support sleeve is arranged outside the kit cover, a movable plate is connected inside the kit cover, the movable plate is provided with a plurality of fastening grooves, the fastening grooves are matched with the test tube covers, and the test tube covers are connected with the fastening grooves.

The invention relates to the technical field of agriculture, in particular to an RNA sequence microRNA-OsmiR5810 for regulating and controlling the grain weight of rice grains, a deletion editor enOgeuIscB-del and application of the RNA sequence microRNA-OsmiR5810 and the deletion editor enOgeuIscB-del. The RNA sequence microRNA-OsmiR5810 for regulating and controlling the grain weight of the rice grains, provided by the invention, is obtained by carrying out RNA extraction and gene cloning on the 12th chromosome of the rice south peduncle 9108 at the position of 7333285-7333425, and the RNA sequence microRNA-OsmiR5810 is in negative regulation and control in the process of regulating and controlling the grain weight of the rice grains; the nucleotide sequence of the RNA sequence comprises: (1) a nucleotide sequence as shown in SEQ ID NO. 1; or (2) a complementary sequence of the nucleotide sequence as shown in SEQ ID NO.1; or (3) a sequence which is at least 90% homologous with the nucleotide sequence as shown in SEQ ID NO.1 and retains the grain weight regulation activity. The microRNA-OsmiRNA5810 for regulating and controlling the grain weight of the rice grains is identified, the regulation and control mode is negative regulation and control, and a new material is provided for high-yield rice breeding.

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.

The utility model provides a tissue grinding tube capable of quickly purifying RNA (Ribonucleic Acid), which relates to the technical field of RNA extraction, and comprises a grinding tube body, an auxiliary grinding piece and a plurality of grinding raised lines, two ends of the grinding tube body are concave inwards to form a grinding cavity, the plurality of grinding raised lines are fixed on the inner wall of the grinding cavity in an annular array, the auxiliary grinding piece is connected with the inner wall of a sealing cover, and the sealing cover is connected with the grinding cavity. When the grinding pipe body is placed in a grinding instrument for use, the auxiliary grinding piece is used for crushing a tissue sample in a grinding cavity during grinding; according to the utility model, during grinding, the movable rod provided with the crushing cutter rotates in an uncertain direction, the crushing cutter can continuously impact a tissue sample with different forces, and by virtue of the high-frequency cutting and crushing actions of the crushing cutter in different directions, the deep disintegration can be carried out on tough fiber tissues, high-toughness meat structures and the like; the internal cell structure and fiber framework are fully destroyed, so that the crushing effect is better realized, and the grinding process is greatly accelerated.

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.

The present application relates to a primer combination for lung cancer detection and its application in lung cancer diagnosis, in particular, a technology for specifically detecting CIZ1b splice body RNA through real-time fluorescent quantitative PCR technology. The method combines blocker sequence to inhibit non-specific amplification of CIZ1a splice body by designing specific primers and probes, and ensures high-specificity detection of CIZ1b splice body RNA. The detection samples include lung cancer tissues and plasma, and cDNA is generated through reverse transcription after RNA extraction of the samples, and real-time fluorescent quantitative analysis is performed. Meanwhile, the method introduces an internal reference gene B2M for standardization correction, and improves the sensitivity and accuracy of the detection. The present application provides a non-invasive and reliable early lung cancer diagnosis tool, which has good clinical application potential.

The invention provides a method for identifying limb regeneration heterogeneity between salamander and xenopus laevis, and relates to the technical field of biology. Comprising the following steps: respectively extracting samples of salamander and xenopus at different stages, and carrying out RNA extraction and high-throughput sequencing on the samples to obtain original data; evaluating the quality of the original data, performing quality control on the original data, and removing low-quality basic groups or linker pollution sequences; constructing a salamander reference genome index, comparing the quality-controlled data with a reference genome, and outputting comparison data; outputting an original count matrix from the comparison data, and calculating a TPM (Trusted Platform Module) value by using the original count matrix in an R language environment; using sequence alignment to extract common genes of the salamander and the xenopus; and performing principal component analysis and correlation analysis on the acquired data. Through transcriptome analysis and sequencing, the molecular mechanism of the limb regeneration capacity heterogeneity of the salamander and the xenopus laevis is analyzed, and guidance is provided for research of other regeneration capacities.

This invention belongs to the field of biological detection technology, and particularly relates to a reproductive tract HPV-mRNA-E6 / E7 detection system and method based on nanopore sequencing. The system includes a sample pretreatment module, an RNA extraction and purification module, a reverse transcription module, a nanopore sequencing module, and a bioinformatics analysis module. The method includes the following steps: (1) reproductive tract sample collection and pretreatment; (2) extraction and purification of HPV-mRNA in the sample; (3) reverse transcription of the purified HPV-mRNA into cDNA; (4) nanopore sequencing of the cDNA to obtain raw sequencing data; (5) filtering, comparing, and annotating the sequencing data through the bioinformatics analysis module to achieve HPV type identification and quantitative analysis of E6 / E7 gene expression levels. This invention utilizes the advantages of nanopore sequencing, such as long read length and real-time detection, combined with the specificity of E6 / E7 mRNA as a biomarker of active infection. It can directly detect latent and active HPV infection, improve detection sensitivity, accurately distinguish infection types, and shorten the detection cycle. It enables rapid, accurate, and high-throughput detection of active HPV infection in the genital tract, providing reliable technical support for early screening, disease monitoring, and prognostic assessment of HPV-related diseases such as cervical cancer.

The present application relates to the field of biotechnology, and in particular to a method and a kit for determining the secondary structure of RNA. The method comprises a sample pretreatment step, an RNA extraction step, a library preparation step and a sequencing step. The library preparation step comprises a reverse transcription step, a first purification step, a second purification step, a linker ligation step, a third purification step, and an amplification and library construction step. The method of the present application has simplified operation steps, low requirements for the initial amount of RNA, and high enrichment efficiency.

A novel plant RNA extraction kit comprises a kit body, a kit body side bag, a first partition plate, a rectangular supporting frame, a sliding buckle, a U-shaped positioning plate, a guide rod, a spring, an upper storage basket, a second partition plate, a kit cover and a rotating supporting plate, the kit body side bag is arranged on the side face of the kit body, the first partition plate is matched with the kit body and inserted into the kit body, and the rectangular supporting frame is arranged on the rectangular supporting frame. The rectangular supporting frame is matched with the kit body, the rectangular supporting frame is inserted into the kit body, the outer side of the rectangular supporting frame is connected with the kit body, the upper storage basket is matched with the kit body, the upper storage basket is inserted into the kit body, the lower surface of the upper storage basket is attached to the rectangular supporting frame, the second partition plate is matched with the upper storage basket, and the second partition plate is inserted into the upper storage basket. The upper surface of the upper storage basket is lower than the upper surface of the kit body.

The invention belongs to the technical field of tumor molecule detection, and particularly discloses a method and a kit for detecting HER2 gene mRNA (messenger Ribonucleic Acid) trace expression based on an RT-ddPCR (Reverse Transcription-Derived Polymerase Chain Reaction) technology. The kit contains a cross HER2 exon 19-20 / 20-21 specific primer probe, a reference gene specific primer probe and reverse transcriptase, and the ddPCR premix solution, the HER2 probe and the reference gene probe are respectively marked with different fluorophores. According to the detection method, after RNA extraction, reverse transcription and RT-ddPCR amplification of a detection sample are carried out, the HER2 / internal reference copy number ratio is calculated to realize typing, a breast cancer sample is completely negative when the Ratio is smaller than 1.0, is ultralow in expression when the Ratio is larger than or equal to 1.0 and smaller than 5.0, is low in expression when the Ratio is larger than or equal to 5.0 and smaller than 50.0, and is medium-high in expression when the Ratio is larger than or equal to 50.0. The detection sensitivity reaches 0.1 copy / microliter, absolute quantification can be achieved, adaptability to FFPE samples is high, breast cancer patients with ultralow expression of HER2 in IHC 0 can be accurately screened, a molecular basis is provided for accurate medication of ADC drugs such as Trastuzumab deruxtecan, and the kit can also be used for related detection of solid tumors such as gastric cancer and cervical cancer with HER2 gene amplification as a marker.

The present application relates to the technical field of biomedical detection, and particularly relates to an intelligent transcriptome analysis system based on a maternal-fetal interface biomarker. The system comprises a maternal-fetal RNA low-temperature storage module, a maternal-fetal genome sequencing calculation module, a transcriptome differential expression determination module and a transcription differential expression network construction module, can obtain a maternal-fetal interface biological blood sample set and perform RNA extraction and low-temperature storage to generate a maternal-fetal blood RNA low-temperature sample group; by constructing a maternal-fetal blood RNA sample library and performing single-end sequencing calculation and reference gene mapping screening, a maternal-fetal RNA reference genome is generated; based on the maternal-fetal RNA reference genome, differential expression genes are determined, target gene clustering and enrichment analysis are simultaneously performed, the gene pathway distribution corresponding to each gene classification is obtained, and gene expression network analysis is performed to generate a maternal-fetal target gene expression network. The present application can analyze the gene pathway network between maternal-fetal target genes.

The invention provides a primer for specifically detecting genome DNA residues in sisal hemp cDNA and a method and application thereof. According to the invention, a beta-ACTIN gene is found in housekeeping genes stably expressed in each tissue of sisal hemp, and a unique sequence difference exists in the beta-ACTIN gene of the sisal hemp by comparing a DNA sequence and a cDNA sequence of the genome. On the basis, a pair of specific primers is developed, cDNA and gDNA of the sisal hemp can be clearly distinguished in common agarose gel electrophoresis on the basis of the length difference of amplified fragments, and gDNA pollution residues can be accurately detected in sisal hemp of different sources and varieties. The detection primer has the advantages of being high in specificity, good in stability, high in universality, simple in detection method, economical, practical and the like, and an efficient and specific standardized quality control tool is provided for evaluating sisal hemp RNA extraction quality and guaranteeing reliability of subsequent molecular experiment data.

The invention discloses a non-toxic RNase inhibition reagent for RNA preservation, the core of the non-toxic RNase inhibition reagent comprises an RNase inhibition component based on an ionic liquid and a natural choline cationic material, the concentration of the component is set to be 3%, exogenous and endogenous RNase with the concentration larger than or equal to 98.7% can be rapidly and efficiently inactivated, and the degradation effect of RNase on RNA is blocked from the source; meanwhile, the reagent has the remarkable advantages of no toxicity and no irritation, completely meets the application standard of a green laboratory, and has good compatibility with mainstream RNA extraction processes of a Trizol method, a column method, a paramagnetic particle method and the like; the reagent is preloaded at a sample collection starting point and carries out immediate protection, so that an RNA sample can be effectively preserved at 4 DEG C or 25 DEG C, and the success rate of RNA extraction is increased by more than or equal to 40%.

The invention belongs to the technical field of biological detection, and particularly relates to a sample pretreatment method for targeted sequencing of high-sensitivity and broad-spectrum respiratory pathogenic microorganisms. The sample pretreatment method comprises the following steps: sequentially carrying out grinding wall breaking and enzymolysis wall breaking on a to-be-detected sample; the grinding and wall breaking time is 4-6 minutes, or / and an RNase inhibitor is added into an initial grinding and wall breaking system. The method can well balance the pathogens difficult to break walls and the pathogens easy to break walls but relatively difficult to stabilize the released nucleic acid in a balanced and high detection rate under the same reaction condition. According to the method, on the premise of ensuring the extraction efficiency of the nucleic acid of the bacteria difficult to break walls, the most suitable mechanical wall breaking time of pathogens easy to break walls but relatively difficult to stabilize the nucleic acid is selected, and the stability and detection of the RNA extraction process are remarkably improved by using the RNase inhibitor.

The invention discloses a single-ended targeting single-ended random molecular tagging method for quantitative analysis of polynucleic acid variation and application of the single-ended targeting single-ended random molecular tagging method. The method comprises the following steps: 1) extracting RNA (Ribonucleic Acid) of a sample; 2) carrying out one-chain reverse transcription by using one-chain reverse transcriptase and a random primer with a joint and UMI; the structure of the random primer from the 5'end to the 3 'end is sequentially as follows: a joint, a 15-base UMI, a 7-base special sequence and a 5-40-base random sequence; 3) synthesizing two-chain cDNA (complementary deoxyribonucleic acid); (4) carrying out first round of nested PCR amplification; (5) carrying out second round of nested PCR amplification; according to the present invention, the specially designed random primers are adopted, the targeting nested PCR is combined, and the capture and the high-specificity amplification of the polynucleic acid variation are simultaneously achieved, and the method is suitable for the high-throughput screening and the dynamic monitoring of the polynucleic acid variation in tumor and other diseases, and provides the reliable technical support for the marker development and the drug effect evaluation.

The invention discloses an RNA co-immunoprecipitation kit for specific binding of prion protein in muscle cells and an RNA extraction method, and relates to the technical field of bioengineering. According to the present invention, the specific co-precipitation kit preparation and the specific extraction steps comprise cell lysis, cell lysis solution-antibody-magnetic bead incubation, protein digestion and RNA extraction, such that the high purity and the sufficient amount of the enriched RNA are successfully ensured so as to completely meet the strict requirements of the subsequent RNA-seq sequencing. The method overcomes the inherent defects of the existing RIP technology in the research of non-classical RNA binding proteins, realizes the optimization of the whole process from high-specificity enrichment to high-quality sequencing sample preparation through the integrated innovation of methodology and the kit, has the outstanding advantages of strong specificity, high sensitivity, good repeatability, simple operation and the like, and has a wide application prospect in the research of non-classical RNA binding proteins. And remarkable technical progress is achieved, and a positive application effect is generated.

The invention discloses a zebrafish cell autophagy rapid detection kit. The kit comprises a rapid RNA extraction column, and a matched lysis solution, a washing solution, an eluent, DNase I, a reverse transcription premix solution, a qPCR premix solution, and a dry powder or premix solution of a specific primer. The invention also discloses an application of the kit. The invention also discloses a method for screening compounds or substances for regulating and controlling zebrafish cell autophagy by using the kit. According to the invention, brand new high-specificity and high-efficiency primers are adopted, so that false positive and background noise are remarkably reduced, and detection data of expression quantities of LC3B and p62 genes are more real and reliable; the optimized integration process greatly shortens the detection time; the premixed reagents and the standardized operation process ensure good reproducibility of experimental results of different batches and different operators.