91 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 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.

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 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 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 primer group for PMAxxTM-qPCR (Polymerase Chain Reaction) of infectious PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) and a detection kit of the primer group. The primer group comprises a pair of primer pairs PRRSV ORF67-UTR '3F / 3R and a fluorescent probe P, the 5'end of the fluorescent probe is marked with a fluorescent reporter group, and the 3 'end of the fluorescent probe is marked with a fluorescent quenching group. The fluorescent quantitative kit comprises the primer group. A one-step method is adopted, the same reverse transcription efficiency is ensured, a trace virus RNA extraction kit is adopted, the stability and reliability of processing results of different batches are ensured, and detection of low-copy virus nucleic acid is ensured.

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.

The invention belongs to the technical field of plant molecular biology, and particularly discloses a diatom RNA extraction method and a method for evaluating influence of pollutants on diatom gene expression. The invention firstly provides a diatom RNA extraction method, RNA is extracted by virtue of a cell homogenate physical crushing method and according to an improved TRIzol method, the release amount of RNA is high, and the purity reaches the standard. The invention also provides a method for evaluating the influence of pollutants on gene expression of diatom, which is based on the diatom RNA extraction method, takes Tubulin as a reference gene, takes one or more of iron redox protein, ferrithionein, ketoacyl coenzyme A thiolase, ethanol dehydrogenase, lipoprotein and acetyl coenzyme A carboxylase as target genes, and evaluates the influence of pollutants on gene expression of diatom. The relative expression level of a target gene is detected through qPCR and can be used for evaluating the acute toxicity of pollutants. The method effectively solves the problems that diatom RNA is difficult to extract, the diatom gene expression level is difficult to detect through qPCR, and the molecular mechanism of pollutant action is difficult to evaluate.

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.