170 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 relates to a method for constructing a plasma or serum exosome RNA quality evaluation model suitable for transcriptome sequencing and application of the method, stably expressed RNA candidate factors are screened, a verified primer probe is used for conducting qPCR quantification on exosome RNA, and a Ct value is obtained; an RNACt value of the exosome is associated with RNA-seq data, a reasonable Ct value is screened and set as a reference threshold value, and the RNA quality of the exosome is graded as a measurement index for judging whether the RNA-seq data is qualified or not, so that the quality of miRNA or mRNA of the exosome can be independently evaluated, and the quality of the two RNAs can also be evaluated at the same time. The method can timely and effectively evaluate whether upstream sample collection, exosome separation, RNA extraction and other methods are appropriate or not, can reduce the link of transferring low-quality samples to a downstream high-throughput library construction experiment, and reduces the cost loss of manpower, samples, reagent consumables and the like.

The invention provides a gene SbERF71 capable of improving waterlogging tolerance of sorghum, protein, application and a breeding method, and belongs to the technical field of crop breeding. On the basis of a sorghum plant with waterlogging tolerance, it is determined that SbERF71 is a key gene affecting the waterlogging tolerance of sorghum through RNA extraction and library building, RNA-Seg data analysis, weighted gene co-expression network analysis (WGCNA) and other methods. The SbERF71 gene is related to gene expression of various stress reaction pathways (such as energy metabolism and aero-tissue formation) in sorghum, is a central regulatory factor of sorghum waterlogging, and is beneficial to reaction of sorghum to flooding stress, expression height up-regulation of sorghum in a flooding stress environment and enhancement of flooding resistance of sorghum, so that sorghum can adapt to waterlogging stress. The invention reveals that sorghum can adapt to molecular and physiological mechanisms of waterlogging stress, provides a new direction for breeding of sorghum waterlogging-resistant varieties, and makes a contribution to food safety in waterlogging agricultural areas.

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.

The invention discloses an RNA fragment extraction method, and relates to the technical field of molecular biology. Comprising the following steps: S1, preparing materials; s2, cell culture: S21, cell resuscitation; s22, performing cell cryopreservation; s23, carrying out cell passage; s3, co-immunoprecipitation, specifically comprising the following steps: S31, binding an antibody to a protein A / G magnetic bead; s32, carrying out cross-linking combination on the antibody; s33, carrying out cell lysis treatment; s34, carrying out manual antigen immunoprecipitation; s4, carrying out RNA extraction on the basis of a TRIZOL method; and S5, carrying out reverse transcription and real-time fluorescent quantitative PCR. By improving the components of the lysis solution and the washing solution, the extraction efficiency and the yield of RNA are improved, and meanwhile, the pollution of impurities is reduced; the usage amount of reagents and experiment steps are reduced, and the experiment cost is reduced.

The invention relates to a kit for diagnosing non-small cell lung cancer as well as a use method and application thereof, and belongs to the field of biological medicines. The kit comprises reagents required by RNA extraction and reverse transcription, and a PCR amplification system containing primers and probes for respectively and specifically targeting SFTPA1 and SFTPA2 genes, wherein the PCR amplification system comprises primers and probes for respectively and specifically targeting SFTPA1 and SFTPA2 genes; the 5'end of the probe is provided with a fluorophore, and the 3 'end of the probe is provided with a quenching agent; the probe and the primer are designed based on specific regions of SFTPA1 and SFTPA2 genes, so that high specificity and high sensitivity of the probe and the primer in gene expression detection are ensured. The use method of the kit comprises the following steps: extracting RNA (Ribonucleic Acid) in a detection sample, reversely transcribing the RNA into cDNA (Complementary Deoxyribose Nucleic Acid), detecting the expression levels of the SFTPA1 and SFTPA2 genes by utilizing a qPCR (Quantitative Polymerase Chain Reaction) technology or a dPCR technology, and finally, preliminarily diagnosing whether a detection object suffers from the non-small cell lung cancer or not by comparing the expression differences of the SFTPA1 and SFTPA2 genes of the detection object and healthy people and non-small cell lung cancer patients. The invention provides a reliable tool for early diagnosis and individualized treatment of the non-small cell lung cancer.

The invention discloses a method for accurately identifying sweet-waxy corn germplasm, and particularly relates to a method for identifying sweet-waxy corn germplasm, which comprises a first tracking process: a process from F3-generation seed budding to F4-generation seed maturation, including sweet-waxy gene DNA tracking marking after seed budding; sampling once 28 days after pollination, and measuring the thickness of soluble sugar, amylose, amylopectin and pericarp; extracting DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) of F3-generation plants in a After the F4-generation seeds are mature and picked, determining sweet-waxy gene DNA, determining amylose and amylopectin, and determining single-plant grain weight, hundred-grain weight and endosperm / embryo dry weight; a second tracking process: a process from germination of the F4-generation seeds to bearing of fresh sweet-waxy corn, including observation and recording of a field corn plant growth process; measuring the thickness of soluble sugar, amylopectin, amylose and pericarp 15 days after pollination; determining and analyzing the traits of the indoor test species; data analysis and germplasm scoring: comparing and analyzing the data obtained in the second tracking process, scoring the F4-generation seeds, and determining the germplasm quality of the F4-generation seeds.

The invention discloses a sesame fusarium wilt identification method based on a molecular marker, and relates to the technical field of phytopathology, the identification method comprises the following steps: S100, RNA extraction and purification: extracting total DNA and protein pollution RNA from a sesame sample to be detected, purifying the extracted RNA by using an efficient RNA extraction kit, removing existing impurities and inhibitors, and determining the content of the protein pollution RNA in the sesame sample to be detected; according to the method, specific long-chain non-coding RNA is adopted as a molecular marker, the molecular marker has high specificity and can accurately distinguish the sesame wilt from other diseases and healthy plants, the probability of misjudgment can be greatly reduced through data analysis, and the detection accuracy is improved. The specific long-chain non-coding RNA has the advantages that the specific long-chain non-coding RNA can be used for detecting the sesame fusarium wilt, reliable guarantee is provided for accurate identification of the sesame fusarium wilt, meanwhile, the specific long-chain non-coding RNA has good stability under different environmental conditions, and detection of a large number of samples can be completed within a short time through PCR amplification and high-throughput sequencing.

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.

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 invention relates to a reagent for high-quality extraction of RNA (Ribonucleic Acid) in mature corn kernels and embryos and an extraction method of the reagent. The reagent is named as RNA extraction auxiliary liquid, starting from the characteristic that corn kernels and embryos are rich in starch, protein, grease, polysaccharide, polyphenol and the like, the reagent for removing impurities in the mature corn kernels and embryos in the RNA extraction process and the extraction method are designed and researched and developed, in the initial stage of extraction, the impurities are removed as much as possible, smooth extraction in the subsequent process is guaranteed, and the extraction efficiency is improved. And finally, high-quality RNA is obtained. The method is suitable for RNA extraction in corn seeds and embryos, and the obtained high-quality RNA can be applied to subsequent PCR (Polymerase Chain Reaction), high-throughput sequencing and other experiments.

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.

Methods of extracting ribonucleic acid from plant tissue involve treating multiple groups of plant seeds with different plant growth regulators. Each group of plant seeds can be treated with a different plant growth regulator. Methods further involve germinating the plant seeds to form seedlings, harvesting tissue samples from subgroups within each group of plant seeds and seedlings at different timepoints, and extracting total RNA from each harvested tissue sample. Methods can also involve sequencing the extracted RNA, determining an expression level of a plurality of genes relative to a negative control, and identifying a subset of differentially expressed genes at each distinct timepoint.

The utility model discloses a front-end treatment device for tumor sample pretreatment and RNA (Ribonucleic Acid) extraction. The front-end treatment device comprises a centrifugal tube and an extraction component, the extraction assembly comprises an extraction pipe group and a liquid injector; the extraction tube group comprises a first tube body, a second tube body and a third tube body, pores are formed in the bottom of the first tube body and the bottom of the second tube body, and a first filter membrane of 20-40 microns is fixedly coated at the bottom of the first tube body; the bottom of the second tube body is coated and fixed with a second filter membrane of 100-300nm; the first pipe body is arranged on the inner side of the second pipe body in a hollow embedded mode, and a first sealing ring is arranged between the upper portion of the outer side of the first pipe body and the upper portion of the inner side of the second pipe body. According to the front-end treatment device for tumor sample pretreatment and RNA extraction, the centrifugal tube is adopted for dewaxing and enzymolysis of a tissue sample, then step-by-step splitting decomposition and filtration are conducted through the extraction assembly, operation is convenient, and the RNA extraction efficiency can be improved.

The invention discloses a molecular mechanism research method regulated and controlled through metabolism reprogramming, and relates to the technical field of molecular biology. The method specifically comprises the following steps: S1, preparing an experimental cell material and an experimental reagent; the experimental cell material is melanoma cells, and the experimental reagent comprises a DMEM high glucose culture medium, PINK1 targeting lentivirus and the like. According to the application, cell infection, RNA (Ribonucleic Acid) extraction, RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction) detection, Western blot experiment, cell activity detection and the like are carried out on lentivirus targeting PINK1, a molecular mechanism of melanoma caused by BRAF gene V600E mutation regulated by PINK1 through metabolic reprogramming is researched, and aerobic glycolysis caused by PINK1 down-regulation can be alleviated by inhibiting ENO2, so that PINK1 inhibits aerobic glycolysis of melanoma cells, and the application has the advantages that the application is simple, the application is convenient, and the application has a wide application prospect. The low-expression PINK1 promotes the growth of melanoma cells.

The invention belongs to the technical field of biology, and particularly relates to an efficient extraction method of mineralized tissue RNA applicable across animal categories. The invention provides a solution combination suitable for mineralized tissue pretreatment, which not only is beneficial to promoting the release of cells from a mineralized matrix, but also can inhibit the activity of ribonuclease to a certain extent, and is beneficial to extracting high-quality RNA (Ribonucleic Acid). Furthermore, the invention also provides an efficient extraction method of mineralized tissue RNA suitable for cross-animal categories, the sample is pretreated by using a solution combination, and then RNA extraction, precipitation and other treatments are sequentially performed, so that the RNA extraction efficiency of the exoskeleton sample is remarkably improved, and the method has good universality and is suitable for large-scale popularization and application. The method is suitable for extracting RNA in cross-animal mineralized tissues, has important biological significance, and lays a methodological foundation for subsequent development of transcriptomics research of exoskeleton dynamic development, analysis of an environmental adaptation mechanism and construction of a biomineralization process regulation and control network.

The invention discloses a preparation method of reusable Tim4-rMGO magnetic beads and application of the reusable Tim4-rMGO magnetic beads in exosome extraction.The preparation method comprises the steps that sterile and enzyme-free water is added into Tim4 protein powder, the Tim4 protein powder is dissolved and then added into an activated carboxylated magnetic graphene oxide solution, after an oscillation reaction, pure water magnetic suction washing is conducted, and the Tim4-rMGO magnetic beads are obtained. The Tim4-rMGO magnetic bead provided by the invention has high selectivity, that is, the Tim4-rMGO magnetic bead only depends on Ca < 2 + > chelation, avoids non-specific destruction of interaction of other proteins or lipids, and can be reused. The eluted exosome does not need an additional desalination step, and can be directly used for RNA extraction, proteomics or function research.

The utility model discloses an RNA (Ribonucleic Acid) extraction kit which comprises a placing assembly, a placing box, a placing block fixed in an inner cavity of the placing box, reagent intubation tubes uniformly fixed in the placing block and a sealing cover hinged on the placing box, and arc-shaped elastic sheets are symmetrically fixed on the inner walls of the reagent intubation tubes; the clamping assembly comprises a threaded rod rotationally installed on the sealing cover, a rotary knob fixed to the outer end of the threaded rod, a guide rod fixed to the sealing cover, a threaded barrel meshed with the threaded rod, a guide barrel fixed to the threaded barrel and connected to the guide rod in a sleeving mode, and a clamping plate fixed to the threaded barrel. The sealing cover is closed, the rotary knob is rotated to drive the threaded rod to rotate, the threaded cylinder and the clamping plate are driven to move and get close to the reagent tube, the top of the reagent tube is abutted, the reagent tube is well fixed, and shaking of the reagent tube is avoided.

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 invention provides a preparation method of a crab cactus X virus CP protein polyclonal antibody, which comprises RNA extraction and RT-PCR, recombinant prokaryotic expression vector construction, ZyVX CP protein prokaryotic expression and protein purification, and ZyVX CP antiserum preparation, and also provides application of the crab cactus X virus CP protein polyclonal antibody for detecting a pitaya ZyVX infected sample. According to the present invention, the CP gene encoded by ZyVX is cloned, the prokaryotic expression vector is constructed, the high immunogenicity ZyVX CP protein is obtained through prokaryotic expression and purification, the specific polyclonal antibody of ZyVX CP is prepared, the rapid Dot-blot-ELISA detection method of ZyVX is established, and the technical scheme is provided for the field prevention and control and research of ZyVX.

The invention discloses a flowering regulating gene of honeysuckle and application thereof, and relates to the technical field of plant gene engineering. The flowering regulating gene of honeysuckle is AGL82, and the gene sequence thereof is shown in SEQ ID NO.1; the amino acid sequence of the protein encoded by the regulating gene is shown in SEQ ID NO.2; the regulating gene can be cloned by RNA extraction and reverse transcription; the regulating gene is used for regulating the flowering period of honeysuckle and for constructing a plant overexpression vector, and the regulating gene can promote the early flowering of honeysuckle; an AGL82 gene related to the flower organ development of honeysuckle and its encoding protein sequence, as well as a method for preparing the AGL82 gene and primers required in the preparation process are provided, and the function of the AGL82 gene in promoting plant flowering is verified by successfully constructing an AGL82 gene overexpression vector, genetically transforming Arabidopsis thaliana and other technical means.

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 relates to a watermelon seed vigor molecular marker and application thereof, the marker is a Cla97C02G040900 gene, and the nucleotide sequence of the marker is as shown in SEQ ID NO: 1. By designing specific forward and reverse primers (SEQ ID NO: 2 and SEQ ID NO: 3), the expression level of the gene in watermelon seeds can be detected. The molecular marker is used for evaluating the vigor condition of watermelon seeds, the expression quantity of the gene is analyzed through real-time fluorescent quantitative PCR, the application comprises the step of identifying the vigor of the watermelon seeds through the watermelon seed vigor molecular marker, and the experimental process of the application comprises the steps of initiation treatment of seed vigor improvement, RNA (Ribonucleic Acid) extraction, cDNA (Complementary Deoxyribose Nucleic Acid) synthesis and PCR detection. Further standard curve making and expression quantity analysis results show that the high expression quantity of the Cla97C02G040900 gene is related to the seed germination rate and activity improvement. The invention provides an accurate and reliable molecular detection tool for watermelon seed vigor evaluation.

The invention relates to the technical field of virus detection, in particular to an SARS-CoV-2 pseudovirus vector construction and verification method which comprises the following steps: selecting target genes based on novel coronavirus, connecting nucleotide sequences of the target genes in series, and cloning the nucleotide sequences to a lentivirus packaging expression vector to obtain plasmids; transforming the plasmid and a second-generation lentivirus packaging plasmid into an escherichia coli cloned strain, carrying out enlarged culture, and extracting an endotoxin-free plasmid; co-transfecting cells with the endotoxin-free plasmids, and collecting pseudovirus particles containing target gene RNA (Ribonucleic Acid) from supernate of the transfected cells; precipitating pseudovirus particles, discarding supernate, and resuspending and concentrating by using a virus dissolving solution; according to the method, a standard curve is drawn for a target gene, RNA extraction, reverse transcription and fluorescent quantitative PCR are performed on pseudovirus particles, the titer of the pseudovirus is calculated through the standard curve, the pseudovirus is obtained, the pseudovirus is not infectious, but the genome of the pseudovirus is similar to that of a real virus, so that the method can be safely used in a laboratory, and the biological safety risk in operation is reduced.

The invention discloses a plasma RNA extraction and library building method for NGS sequencing and application of the plasma RNA extraction and library building method, a developed rRNA removal reagent and a DNA stop buffer are used for an rRNA removal step and a DNA removal step in a library building process, so that the problem of sample loss during DNA removal in the prior art and the problem of existing mt-rRNA removal cost are effectively solved; the rRNA proportion in the library is reduced, so that the universality of NGS sequencing in RNA sequencing is improved, and the possibility is provided for large-scale sequencing.