200 results about "Total rna" patented technology

The invention discloses a verification method for regulating and controlling proliferation and differentiation of bovine adipocytes by gene interference, which comprises the following steps: cutting bovine adipocyte tissues into small blocks, directly pasting the small blocks in a culture dish for culture, and promoting cell growth and proliferation by using nutrition and growth factors of the tissues; a ZNF423 gene is targeted by adopting an siRNA technology, siRNA is transfected into bovine fat cells by utilizing a transfection reagent, and the siRNA is an interference fragment containing si-ZNF423; the method comprises the following steps: respectively extracting total RNA (Ribonucleic Acid) and total protein of bovine adipocytes, and detecting by utilizing qPCR (Quantitative Polymerase Chain Reaction) quantitative analysis, a WB (White Batch) technology, EdU dyeing, flow cytometry detection, oil red O, BODIPY (Boron Dipyrromethene), Nile Red dyeing, TG (Triglyceride) content determination and the like. According to the invention, ZNF423 gene expression is interfered by the interference fragment containing si-ZNF423, and the effect of inhibiting proliferation and differentiation of bovine adipocytes is detected by various methods, so that the effect of the interference fragment in the bovine adipocytes is known conveniently, and regulation and control of proliferation and differentiation of the bovine adipocytes are realized by interfering gene expression by the interference fragment. The beef quality can be improved; the beef cattle breeding can be optimized.

The invention belongs to the technical field of biology, and particularly relates to a reagent combination and a kit for extracting nucleic acid through a paramagnetic particle method and application of the reagent combination and the kit. The reagent combination or the kit does not adopt beta-mercaptoethanol with pungent smell or dithiothreitol needing to be stored at low temperature as an antioxidant, but adopts tris (2-formylethyl) phosphine hydrochloride and / or metabisulfite as an antioxidant, belongs to common salt chemical reagent powder, can be stored at normal temperature, and can be used for preparing a reagent for the first time. Manual addition by experimenters before an experiment is not needed, so that nucleic acid (RNA) released by splitting decomposition can be effectively prevented from being oxidized and denatured, and the integrity of extracted nucleic acid (total RNA) fragments is ensured; especially, the metabisulfite is commonly used in food additives, does not cause harm to the body, and is non-toxic and non-volatile.

The invention discloses a gastric signet-ring cell carcinoma biomarker and a detection kit thereof, belongs to the technical field of biological medicines, and particularly relates to the detection kit of the gastric signet-ring cell carcinoma biomarker, which comprises a total RNA (Ribonucleic Acid) extraction reagent, an RNA purification reagent, a cDNA (Complementary Deoxyribonucleic Acid) first strand synthesis premixing reagent, a rapid fluorescent quantitative premixing reagent (SYBR Green), nuclease-free double distilled water and a specific primer. The RNA purification reagent at least comprises isopropanol, chloroform and absolute ethyl alcohol; the specific primer comprises an upstream specific primer and a downstream specific primer. The detection kit for the gastric signet-ring cell carcinoma biomarker is high in RNA purification and recovery efficiency and excellent in specificity.

The invention relates to the technical field of molecular biology, in particular to a fluorescent quantitative PCR reference gene screening method in different tissues and stress treatment of perilla frutescens, which specifically comprises the following steps: step 1, respectively selecting perilla frutescens leaves, stalks, roots, fruits at different development stages after flowering and perilla frutescens leaves treated by 20% PEG6000 for 0H, 3H, 6H, 9H, 12H and 24H as samples; extracting total RNA (Ribonucleic Acid) of each sample, and carrying out reverse transcription on the RNA to synthesize cDNA (Complementary Deoxyribose Nucleic Acid) as And step 2, screening out candidate reference genes of the purple perilla by using the sequencing data of the early-stage purple perilla transcriptome, namely PfActin, Pf18S, PfEF-1alpha, PfCYP, PfTUA, PfTUB and PfGAPDH respectively, and screening out the candidate reference genes of the purple perilla, namely PfActin, Pf18S, PfEF-1alpha, PfCYP, PfTUA, PfTUB and PfGAPDH. According to the method, the most stable reference genes in perilla leaves, stems, roots, fruits in three development stages and perilla leaves treated by 20% PEG6000 for 0H, 3H, 6H, 9H, 12H and 24H are screened from seven candidate reference genes, and a foundation is laid for deep excavation of functional genes in the later period.

The invention relates to the technical field of drug resistance detection, in particular to a method for detecting cis-platinum drug resistance of bladder cancer based on an exosome miRNA marker and application, and the method comprises the following steps: (1) extracting an exosome from serum of a bladder cancer patient; (2) extracting total RNA (Ribonucleic Acid) from the exosome; (3) detecting the expression level of specific miRNA in the total RNA; (4) on the basis of the expression level of the miRNA, the drug resistance risk of a patient to cis-platinum is evaluated, and the prediction accuracy (AUC is increased from 0.72 to 0.89) of the cis-platinum drug resistance of bladder cancer is remarkably improved; compared with the prior art, the treatment effective rate is greatly improved (from 45% to 60%), meanwhile, the occurrence rate of serious adverse reactions is reduced (from 25% to 15%), and in a drug-resistant bladder cancer model, the comprehensive scheme shows that the tumor inhibition rate exceeds 70% and is far higher than that of a traditional method.

The invention provides a macro virus group sequencing library construction method for removing host ribosome RNA through an enzyme method and a kit, and belongs to the technical field of biology. The method comprises the following steps: hybridizing total RNA (Ribonucleic Acid) of a sample with a DNA (Deoxyribose Nucleic Acid) probe aiming at host ribosome RNA to form a DNA / RNA hybrid chain; rNase H is used for digesting RNA in the hybrid chain, and DNase I is used for digesting the remaining DNA probe in the system; purifying to obtain virus RNA (Ribonucleic Acid); carrying out fragmentation treatment on the virus RNA; carrying out reverse transcription on the fragmented RNA to synthesize a first chain cDNA, then synthesizing a second chain cDNA, and carrying out terminal repair and dA tail addition to obtain double-chain cDNA; connecting the linker with the dT tail with the double-chain cDNA; and carrying out PCR amplification and purification on the connection product to obtain the macro virus group sequencing library. According to the construction method and the kit, the problem that the full-length sequence of the virus cannot be obtained from a low-load sample can be solved.

The invention provides a method for eliminating ribosomal RNA in a mouse total RNA sample, which comprises the following steps: carrying out reverse transcription on the total RNA in the mouse sample into cDNA, and amplifying the cDNA by adopting a primer sequence group to obtain a DNA fragment covering all sequences of the mouse ribosomal RNA; carrying out amplification by taking the DNA fragment as a template and adopting a single primer in the primer sequence group to obtain a single-stranded DNA probe; hybridizing the total RNA of the mouse sample by adopting a single-stranded DNA probe, and performing RNase H digestion to obtain a digestion product; sequentially carrying out DNase I digestion and RNA purification on the digestion product to obtain a target product; wherein the primer sequence group comprises upstream primers from the first to the nineteenth and downstream primers from the first to the nineteenth, and the primer sequence group has the characteristics of low cost, stable performance and high reduction efficiency.

The invention discloses an application method for revealing RIPK3 as a pathological index of osteoarthritis based on transcriptomics analysis, and belongs to the technical field of molecular biology and genetic engineering.The method comprises the following steps that S1, a Trizol reagent is used for conducting total RNA extraction on an articular cartilage specimen, and the RIN number gt is selected; 7.0, constructing a sequencing library by using the high-quality RNA sample; s2, carrying out quality inspection on the constructed library, and carrying out mRNA sequencing by using an Illumina NovaseqTM6000 sequencer after all the libraries are qualified in quality inspection; s3, carrying out bioinformatics analysis on data generated by high-throughput sequencing; s4, different parts of the cartilage tissue of the OA patient are labeled with RIPK3 through immunofluorescence, it is proved that expression of the RIPK3 in cartilage cells has strong correlation with the pathology of the OA focus, and the feasibility of taking the RIPK3 as a clinical pathology index is proved.

The invention belongs to the technical field of pseudorabies virus prevention and control, and discloses application of BAY87-2243 in preparation of a drug for inhibiting pseudorabies virus proliferation, and the application is as follows: firstly, PK-15 is infected with a PRV-GD-WH strain, total RNA of cells is taken, and cDNA is obtained through reverse transcription; the change of the transcriptional level of a glycolysis key gene in a cell is detected through fluorescent quantitative PCR (Polymerase Chain Reaction) of an SYBR Green method. BAY87 is further used for acting on the HIF-1alpha, the regulation and control effect of the HIF-1alpha on glycolysis is verified, and finally, the virus load in cells is detected through SYBR Green fluorescent quantitative PCR (Polymerase Chain Reaction) and the inhibition effect of BAY87-2243 on PRV proliferation is determined through TCID50. Meanwhile, the invention also provides a pseudorabies virus inhibition drug.

The present invention discloses a kit for detecting residual total RNA of Escherichia coli, a detection method and an application thereof. By designing a pre-treatment system, a plasmid sample can be directly used for quantitative detection of residual total RNA of Escherichia coli by RT-qPCR after digestion. The detection method eliminates the influence of the enzyme digestion process on the detection result by setting a digestion quality control product, ensures the accuracy of the entire detection result, and has excellent specificity, accuracy, sensitivity and precision.

The invention discloses a nano antibody for resisting a listeria monocytogenes MurA protein and application of the nano antibody, and relates to the technical field of biology. The amino acid sequence of the nano antibody is as shown in SEQ ID NO.3. MurA protein of prokaryotically expressed listeria monocytogenes is used as an immunogen to immunize alpaca, total RNA of peripheral blood lymphocytes is extracted, VHH fragments are amplified through reverse transcription and two rounds of nested PCR, the VHH fragments and a pComb3Xss vector are converted into TG1 competent cells after enzyme digestion and connection, and a phage display technology is used for three rounds of elutriation, so that the total RNA of the listeria monocytogenes is obtained. The nanometer antibody for resisting the listeria monocytogenes MurA protein is successfully screened out. The nano antibody can specifically recognize MurA protein and whole listeria monocytogenes, and the immunocolorimetric sensor based on thiolated bacteriophage induced colloidal gold aggregation, which is established by utilizing the nano antibody, is strong in specificity, high in sensitivity and low in detection limit, and can greatly shorten the detection time.

The invention provides a construction method and an application method of a wheat yeast two-hybrid cDNA library, and belongs to the technical field of biology. The invention provides a construction method of a yeast two-hybrid cDNA library, which comprises the following steps: extracting total RNA, separating mRNA, converting into cDNA, transferring into escherichia coli competent cells through a series of reactions to obtain a cDNA primary library bacterial liquid, separating primary library plasmids, and obtaining a yeast two-hybrid cDNA library bacterial liquid through an LR recombination reaction and conversion of the escherichia coli competent cells. A high-quality wheat variety Zheng wheat 158 is taken as a research material, wheat gliadin genes Tagli-gamma-1 and Tagli-gamma-11 are cloned, a corresponding bait vector is constructed, the interaction protein is screened from a cDNA library of the Zheng wheat 158 by utilizing a yeast two-hybrid technology, and a foundation is laid for carrying out interaction protein screening of genes related to wheat quality and analyzing a regulation mechanism of the interaction protein in a later period.

The invention relates to the technical field of biological research data analysis, and discloses a computational biological research data analysis display method, which comprises the following steps of: extracting total RNA (Ribonucleic Acid) from biological tissues or cells, reversely transcribing the RNA into cDNA (Complementary Deoxyribonucleic Acid), constructing a library, and sequencing the library by utilizing a high-throughput sequencing platform to obtain a large number of short sequences. According to the method, the number of the short sequences mapped to the same genome coordinate interval is compared and the threshold value is judged, so that the abnormal repeated area can be effectively identified, whether the gene expression difference result is credible or not is clearly judged, and when the number of the abnormal sequences exceeds the preset repeated sequence threshold value, the influence operation of the repeated sequences is eliminated, and the accuracy of the gene expression difference result is improved. That is to say, completely identical and overlapped short sequences are removed under the condition that the FPKM value is relatively high, and the updating result is compared again, so that the problem of expression quantity distortion caused by repeated reckoning can be corrected.

The invention discloses a molecular marker of resistance of chlamydomonas reinhardtii to diflufenican and application of the molecular marker to classification of sensitive populations and resistant populations of the chlamydomonas reinhardtii to the diflufenican, and the molecular marker is CrPDSA113V mutation or CrPDSAF269Y mutation, namely Ala-1113-Val mutation or Ph-269-Tyr mutation. The detection method comprises the following steps: firstly, extracting total DNA and total RNA of chlamydomonas reinhardtii to be detected, taking the total DNA and CDNA after reverse transcription as templates, carrying out PCR (Polymerase Chain Reaction) amplification by using the specific primer pair to obtain PDS gene segments of an amplification product, analyzing whether the PCR amplification product contains the molecular marker or not, and if the PCR amplification product contains the molecular marker, judging whether the PCR amplification product contains the molecular marker or not. Whether the chlamydomonas reinhardtii to be detected is a diflufenican sensitive population or a diflufenican resistant population can be judged. The molecular marker of the resistance of the chlamydomonas reinhardtii to the diflufenican is determined for the first time, and a rapid molecular detection means is provided for the resistance of the chlamydomonas reinhardtii to the diflufenican.

The present invention relates to an RT-qPCR detection method and application for the distribution of human mesenchymal stem cells in mice, belonging to the field of biomedicine technology. The RT-qPCR detection method comprises the following steps: sampling: taking organ tissues of mice injected with human mesenchymal stem cells, extracting their total RNA for testing; detection: using real-time fluorescence quantitative PCR to detect the human mesenchymal stem cell-specific protein gene sequence in the above-mentioned total RNA to obtain the distribution of human mesenchymal stem cells in mice. The above-mentioned detection method can achieve the purpose of quickly and accurately detecting the in vivo distribution of multiple tissues at different time points, providing data support for the selection of stem cell products, and being used in the evaluation of the distribution of stem cells in the body.

The invention discloses a rapid sterile detection method and kit based on a 16S / 18S rRNA-cDNA digital PCR technology, and the method comprises the following steps: S1, extracting total RNA in a to-be-detected sample to obtain bacteria 16S rRNA and fungi 18S rRNA; s2, carrying out reverse transcription on the bacterium 16S rRNA and the fungus 18S rRNA to obtain a stable cDNA (complementary deoxyribonucleic acid) fragment; and S3, carrying out digital PCR amplification by taking the cDNA fragment as a template, and judging whether bacterial contamination exists or not according to an amplification result so as to realize rapid sterile detection. According to the characteristics of high conservative and high copy of bacteria 16S rRNA / fungi 18S rRNA, 30 sets of universal primer probe combinations are verified, and gt can be covered by selecting the optimal two sets of primer probe combinations; 99.99% of common pollution bacteria / fungi in clinic and pharmacy, and no cross amplification with mitochondrial rRNA of mammals such as human, hamster, cattle and the like.

The invention belongs to the technical field of plant genetic engineering, and particularly discloses a nitrate ion transport channel gene PbrSLAH3b and application of the nitrate ion transport channel gene PbrSLAH3b in promoting growth of pear pollen tubules. The method comprises the following steps: extracting total RNA (Ribonucleic Acid) of Dangshan pears from pollen of Dangshan pears; carrying out reverse transcription on the RNA to obtain cDNA (complementary deoxyribonucleic acid); and by taking the cDNA as a template, carrying out amplification to obtain the pear pollen tube nitrate transport channel gene PbrSLAH3b, the PbrSLAH3b gene provided by the invention belongs to a slow anion channel SLAC / SLAH family member, the PbrSLAH3b is remarkably up-regulated in expression in the early growth stage of a pollen tube, and the PbrSLAH3b is positioned in a plasma membrane. The invention verifies that PbrSLAH3b can form a functional anion channel with NO3 <-> selectivity under the condition of not depending on proton gradient or phosphorylation state, and the PbrSLAH3b has relatively strong selectivity on NO3 <->. Therefore, the PbrSLAH3b gene can mediate the absorption and transport of the pollen tube to NO3 <-> and maintain the polarity growth directivity of the pollen tube, and has important application value for revealing the ion regulation mechanism of the growth of the fruit tree pollen tube and improving the reproductive development of the fruit tree.

The invention relates to a method for preparing a TCR-T targeting SARS-Cov-2 and its application, comprising: 1) synthesizing a SARS-Cov-2 antigen peptide XG2; 2) stimulating the peripheral blood of volunteers with HLA-A*02:01 with antigen peptides to obtain SARS-Cov-2 specific T cells; 3) extracting total RNA of SARS-Cov-2 specific T cells, reverse transcribing to obtain cDNA, and performing NSG sequencing after multiple PCR to obtain an immune repertoire; 4) performing bioinformatics analysis on the immune repertoire to obtain a specific CDR3 sequence; 5) using the specific CDR3 sequence to clone and construct a TCR-T lentiviral vector, which is used to target and identify cells infected with SARS-Cov-2 and to kill them.

The invention provides a fish creatine kinase sensitization and immune cross reactivity analysis method which comprises the following steps: preparing a fish muscle water-soluble protein crude extract, separating 40-45kDa protein components in the fish muscle crude extract through SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis), and screening a target sensitization protein combined with IgE / IgG through immunoblotting; the method comprises the following steps: extracting total RNA of grass carp and synthesizing cDNA, designing a creatine kinase specific primer for amplification to obtain a creatine kinase gene sequence, and carrying out bioinformatics analysis after sequencing verification; the method comprises the following steps: cloning a creatine kinase gene to an expression vector, transforming escherichia coli BL21, performing IPTG induced expression, and performing urea gradient dialysis renaturation on an inclusion body to obtain the recombinant grass carp creatine kinase. Detecting the IgE binding activity of the recombinant grass carp creatine kinase and the serum of the allergic patient through dot hybridization; the IgG cross reactivity of the recombinant grass carp creatine kinase and the blue crab arginine kinase is verified through indirect ELISA; the IgE cross reaction degree of the recombined grass carp creatine kinase and the blue crab arginine kinase is quantified through inhibitory dot hybridization.

The invention relates to the field of crayfish evaluation, and discloses a method and a system for evaluating the WSSV resistance of procambarus clarkii, and the method comprises the following steps: sampling the procambarus clarkii, extracting target tissues from a sample, and extracting total RNA (Ribonucleic Acid) for accurate quantification of concentration and synthesis of cDNA (Complementary Deoxyribonucleic Acid); then TRIM23 gene quantitative detection is carried out on procambarus clarkii through a probe method, the purpose of calculating the Ct value is achieved, and finally the WSSV resistance of the procambarus clarkii is evaluated in combination with a relative quantitative mathematical model. According to the method, high pathogenicity or virus operation is avoided, the safety of the operation process is improved, the monitoring process is greatly shortened, and the accuracy is improved.

The application relates to the field of biomedical technology, in particular to a heavy chain and light chain variable region gene sequence of a LSDV ORF123 monoclonal antibody, an expression immunoglobulin full gene sequence and application thereof, the application extracts total RNA of hybridoma cells (1G1-1G12) by hybridoma sequencing, takes mRNA in the total RNA as a template, carries out reverse transcription to obtain cDNA of corresponding antibody genes, and then uses specific PCR to obtain a heavy chain variable region (Variable region of heavy chain, V H ) and a light chain variable region (Variable region of light chain, V L ) of the corresponding antibody, and carries out cloning and sequencing analysis. Finally, the heavy chain and light chain variable region gene nucleotide sequences of a monoclonal antibody (1G1-1G12) targeting the LSDV ORF123 protein are obtained, and the application is realized. Sequence analysis of the heavy chain (V H ) and light chain antibody (V L ) of the above-mentioned monoclonal antibody shows that the V H and V L gene subtypes of 1G1-1G12 are as follows: the heavy chain is IgG type, and the light chain is kappa type.

The invention relates to the field of plant extracts, in particular to a torreya grandis total RNA extract which at least comprises mRNA, tRNA, miRNA and tsRNA. The torreya grandis total RNA extract can be used as a natural active matter for an external preparation for the skin, and the effects of resisting oxidation, resisting aging or resisting wrinkles and tightening are achieved, so that the skin aging is delayed.

The invention belongs to the technical field of gene interference, and discloses a coral larva RNAi gene editing method and application thereof, and the coral larva RNAi gene editing method comprises the following steps: S1, soaking coral larvae in filtered seawater; s2, an RNAi reagent is added, and siRNA transfection is carried out; and S3, extracting total RNA from the transfected coral larvae, carrying out reverse transcription to obtain cDNA, and carrying out qPCR detection on the cDNA. The method comprises a double-stranded siRNA agent formed by a sense strand and an antisense strand aiming at coral calcavity protein, a transfection method and a qPCR (quantitative polymerase chain reaction) detection method. According to the successfully constructed RNAi-based coral cornu cervi larva gene editing method, functional verification of a target gene can be realized, and a powerful tool is provided for analyzing a coral biological mechanism.

The present invention discloses an AfGF14i gene of Aechmea fasciata, a cloning method, an expression vector and applications thereof, belonging to the field of biotechnology. Specifically, total RNA of Aechmea fasciata is extracted, reverse transcribed into cDNA, and taking this as a template, specific primers are designed, and the full-length CDS of the AfGF14i gene is obtained by PCR method, ligated with a vector, and transformed into Escherichia coli Trans-T1 competent cells, and positive clones are picked to obtain the AfGF14i gene of Aechmea fasciata. The full-length CDS of this gene is 729 bp, encoding a protein containing 243 amino acid residues. The present invention uses transient expression and fluorescence microscopy to detect that the subcellular localization of the protein encoded by the AfGF14i gene is the cell membrane. Through transgenic technology, it is found that the overexpression of the AfGF14i gene promotes the early flowering of Arabidopsis thaliana. The acquisition of the AfGF14i gene lays a foundation for studying the flowering mechanism of Aechmea fasciata, provides a theoretical reference for overcoming the high-temperature obstacle of ethylene-induced flowering of pineapple in production, and has important theoretical and practical significance.

This invention discloses a method for constructing a tomato core membrane-associated protein interaction library based on Gateway technology under Fusarium oxysporum infection conditions and its application, relating to the field of genetic engineering technology. The method includes: using tomato root tissue infected with Fusarium oxysporum as material, extracting total RNA and enriching mRNA; synthesizing first-strand cDNA using Biotin-attB2-Oligo(dT) primers; synthesizing double-stranded cDNA enzymatically; ligating it with a specific double-stranded attB1 adapter; introducing attB1 and attB2 sites at both ends of the cDNA; cloning the cDNA into the donor vector pDONR222 via BP recombination to construct a primary entry library; and recombinating the primary entry library with vectors pGADT7 and pPR3-N via LR recombination to construct a nuclear secondary library and a membrane secondary library, respectively. The constructed libraries have high titers, long insert fragments, and low empty vector rates. This invention also relates to the application of the library in screening the interaction between Fusarium oxysporum effector proteins and tomato nuclear membrane-associated proteins, providing an efficient and specific resource platform for studying plant-pathogen interaction mechanisms.

A method for detecting and treating Type II Diabetes includes the use of hsa_piR_020485, a PIWI-interacting RNA isolated from urinary extracellular vesicles (ECVs), as a biomarker for the diagnosis and treatment of Type 2 diabetes mellitus (T2DM). The method is non-invasive, utilizing the differential expression of hsa_piR_020485 in diabetic versus non-diabetic subjects to identify subjects in need of treatment for T2DM. The method includes obtaining a urine sample from a subject, isolating urinary ECVs from the urine sample, extracting total RNA from the isolated urinary ECVs, quantifying hsa_piR_020485 levels from the total RNA, determining if the hsa_piR_020485 expression level exceeds a threshold, and administering one or more T2DM treatments.

The invention belongs to the technical field of molecular biology, and particularly relates to an anti-GLP-1R nano antibody as well as a preparation method and application thereof. The invention provides an anti-GLP-1R nano antibody, and also provides a construction method of the anti-GLP-1R nano antibody. The method comprises the following steps: constructing GLP-1R overexpression plasmids and overexpression cell strains, immunizing alpaca by using the plasmids and the cells to obtain PBMC cells, extracting total RNA, carrying out reverse transcription to obtain cDNA, amplifying antibody gene segments through PCR, connecting the segments to phage expression vectors, and electrically transforming the constructed vectors into SS320 competent cells to finally obtain the GLP-1R immune antibody library. The method comprises the following steps: carrying out cell panning for one round, carrying out Cell ELISA detection to obtain an antibody combined with GLP-1R, carrying out sequencing to obtain an antibody sequence, obtaining antibody protein through mammalian cells, and carrying out FACS detection by using CHOK1-GLP-1R cells to finally obtain the antibody combined with GLP-1R. The method is simple and convenient to operate, and the activity of the antibody obtained by using a mammalian cell expression system is closer to that of a natural antibody.

The invention relates to the technical field of biological pharmacy, in particular to a human serum albumin targeted nano antibody and application thereof. An alpaca is immunized by using an HSA protein, cDNA is obtained through reverse transcription of total RNA of peripheral blood lymphocytes to serve as a template to amplify a nano antibody sequence, finally, five nano antibodies are obtained through separation and are named as antiHSA-A6, antiHSA-A7, antiHSA-B1, antiHSA-E8 and antiHSA-H7 respectively, the five nano antibodies have different antigen complementarity determining regions, and an ELISA result shows that the five nano antibodies have different antigen complementarity determining regions. And the five nano antibodies all show high-affinity binding with human HSA (Human Serum Albumin) protein. The nano antibody provided by the invention is expected to be used for preparing vaccines for treating and / or diagnosing diseases related to HSA, or for prolonging the half-life period of macromolecular drugs in vivo, and has important significance.

The present application relates to the technical field of biological research data analysis, and discloses a kind of computing biology research data analysis display method, including extracting total RNA from biological tissue or cell, RNA reverse transcription is cDNA, constructs library, and utilizes high-throughput sequencing platform to sequence library, to obtain a large number of short sequences.The present application can effectively identify abnormal repeat regions by comparing and threshold judging the number of short sequences mapped to the same genomic coordinate interval, so as to determine whether the gene expression difference result is reliable, when the number of abnormal sequences exceeds the preset repeat sequence threshold, by eliminating the influence of repeat sequence operation, i.e. for the case of high FPKM value, remove the completely same and overlapping short sequences, and re-align the updated results, the expression distortion problem caused by repeat counting can be corrected.

The invention provides a primer group for detecting a tomato yellow mottle related virus TYMaV based on RT-RAA-CRISPR / Cas12a. The primer group comprises an RT-RAA amplification primer pair and a CRISPR / Cas12a detection probe. The invention provides a method for visually and rapidly detecting TYMaV. The method comprises the following steps: extracting total RNA (Ribonucleic Acid) of tobacco leaves infected by TYMaV or coarsely extracted virions; the method comprises the following steps: preparing RT-RAA reaction mixed liquid at the bottom of a PCR tube and CRISPR premixed liquid at the cover of the PCR tube, adding total RNA or crude extraction virus particles into the RT-RAA reaction mixed liquid, incubating in a 42 DEG C metal bath for 30 minutes, placing the CRISPR premixed liquid at the bottom of the tube through centrifugation or shaking, mixing with an RT-RAA reaction product, incubating in a 37 DEG C metal bath for 20 minutes, adding 80 mu L of ddH2O, and detecting a target strip by using a flow measurement chromatography test strip. The method can be used for preparing a kit for detecting TYMaV. According to the primer group and the established TYMaV visual rapid detection method based on RT-RAA-CRISPR / Cas12a, a theoretical basis and a technical guarantee are provided for monitoring, predicting, forecasting, preventing and treating the TYMaV in the field.