48 results about "Trizol" patented technology

The invention provides a method for separating exosomes from animal plasma and for detecting purity. The method mainly comprises the following steps: first, thinning the plasma by virtue of a PBS (phosphate buffer solution) at the ratio of 1 to 1; removing cell components and cell debris in the plasma through centrifuging; precipitating the exosomes through ultrahigh-speed centrifugation; suspending the exosomes by virtue of 250[mu]l of the PBS solution; extracting total RNA in an exosome suspension by virtue of Trizol LS; and conducting poly (A) reversing and related gene quantification on the extracted total RNA, and detecting the purity. The method disclosed by the invention is conducive to researches on plasma exosome contents and exosome related functions in the future.

The invention relates to a detection kit for detecting the relative expression level of PML-RAR (alpha) fusion gene by use of fluorescent quantitative PCR (polymerase chain reaction) technology. The detection kit comprises the following materials in part ratio: red blood cell lysis buffer, TRIzol, chloroform, absolute ethyl alcohol, reverse transcription PCR reagent, detection system PCR reaction liquid, positive control substance and negative control substance. The kit provided by the invention has the advantages that the precision is high, the result is convenient to read, both the amplification efficiency and the rate reach the best level and the like; the complicated condition grope link is omitted; and the experimental efficiency is greatly improved. The kit has good specificity and high sensitivity and is easy and convenient to operate according to the test. The detection kit is favorable for the trace residue detection of the PML-PAR (alpha) (type L / type S) fusion gene in a clinical APL (acute promyelocytic leukemia) patient, and has great importance in performing timely intervention therapy to avoid hematological recurrence, adjusting the therapeutic schedule, evaluating the therapeutic effect, predicting the prognosis and preventing clinical recurrence.

The invention belongs to the technical field of biology and discloses an extraction method of plant RNA (ribonucleic acid). The method comprises the following steps: sufficiently grinding the plant material in liquid nitrogen to powder, adding an extraction buffer A, sufficiently reacting, adding an extraction buffer B, evenly mixing to react, precipitating with isopropanol, separating out the RNA, washing with ethanol, and dissolving in RNase-free water. The method does not need to use DEPC or the expensive extraction kit, and does not need to use phenol, chloroform and other toxic irritating reagents; and the method is simple to operate and easy to implement, has short extraction time (not more than 30 minutes for the whole process) and has the advantages of favorable completeness and higher concentration as compared with the extracted RNA and kit. The method is more suitable for extracting RNA from polyphenol plants from which TRIZOL can not be extracted generally. The purity and concentration of the extracted RNA are sufficient for RNA reverse transcription, RT-PCR (reverse transcription-polymerase chain reaction) and Nothern blot hybrid experiments.

The method provides a method for extracting total ribonucleic acid (RNA) of fish brain tissue. The method specifically includes subjecting the fish brain tissue to liquid nitrogen quick freezing, trizol lysate treatment, low-temperature high-speed centrifugation, and extraction by ammonium sulfate and chloroform n-butyl alcohol, adding isopropanol-sodium acetate for mixing, subjecting a mixture to adsorption and purification through an RNA purification and adsorption column, performing deoxyribonucleic acid (DNA) enzyme treatment, and performing washing to obtain total RNA of the fish brain tissue. According to the method for extracting the total RNA of the fish brain tissue, the influence of lipids (such as phospholipid, glycolipid, and cholesterol) and proteins in the fish brain tissue on RNA extraction is avoided, and the RNA separation and purification method is stable in purity, good in integrity, high in repeatability, and free of pollution.

The invention discloses a method for extracting and separating total proteins of mice sperms by using two-dimensional electrophoresis. The method comprises the following steps: preparing sperms, precipitating DNAs of the sperms after lysis of the sperms, preparing sperm proteins, preparing a protein extraction sample, carrying out first-dimensional isoelectric focusing electrophoresis, carrying out second-dimensional SDS-polyacrylamide gel electrophoresis, dyeing by using coomassie brilliant blue, preparing gel and scanning by using an image scanner. According to the method, lysis of the sperms is completely performed by Trizol, the proteins are extracted by chloroform, impurities are removed by isopropanol, guanidine hydrochloride and ethyl alcohol, and the proteins are purified by acetone, so that the problems of low content of the mice sperms, collection difficulty, small cell volume, a small amount of cytoplasm and a large amount of acrosomal protease are well solved; the sample prepared by the method is complete to prepare, has a large quantity of protein isolating points, and is high in repeatability and clear in chromatogram; by virtue of detection of PDQuestsoftware, the number of the protein points is greater than 1,000; therefore, the method lays the foundation for further mass spectrometry.

The invention discloses an extraction method for effectively enriching tiny RNA in serum or plasma, which comprises the following steps: preseparating the tiny RNA in a sample with Trizol and chloroform, capturing the preseparated tiny RNA in the sample by a method of magnetic beads marked with -Si-OH functional groups, separating the magnetic beads from the sample by a magnetic separator, washing and drying simply, adding RNA-free enzyme water, heating and eluting by a water bath, and further separating by the magnetic separator. The method of the invention can be used for the large-scale verification of the tiny RNA which is a candidate disease maker and the extracting of the tiny RNA during clinical practices, can efficiently enrich the tiny RNA in serum or plasma, is convenient and fast and has low price.

The invention relates to the technical field of biology, in particular to a method for separation and extraction of DNA and RNA in cells. The method includes: a) adopting Trizol for splitting a cell sample to release nucleic acid to obtain a split product; b) contacting the split product with at least one organic extracting solvent, and centrifuging to layer the split product into an upper RNA aqueous phase layer, a middle DNA flocculent layer and a lower organic phase layer; c) respectively subjecting the RNA aqueous phase layer and the DNA flocculent layer to precipitation and washing, wherein a DNA precipitation agent for precipitation comprises 1,2-propanediol. By adoption of the method, problems of low yield and poor purity in simultaneous extraction of DNA and RNA can be effectivelyavoided.

The invention discloses a method for extracting total RNA of the intermuscular bone of megalobrama amblycephala, which comprises the following steps: 1) with megalobrama amblycephala as the material, fetching the intermuscular bone thereof and putting into a sterilized and pre-cooled mortar, and grinding until the intermuscular bone is ground into powder; transferring the powder into a centrifugal tube; adding Trizol and shaking uniformly; and placing at room temperature; 2) centrifuging at 4 DEG C, fetching the supernate, adding chloroform the volume of which is 1 / 5 that of the supernate, shaking so that the mixture is sufficiently emulsified into a uniform white emulsion without layering, and standing at room temperature; 3) centrifuging at 4 DEG C, fetching the supernate and repeatedly extracting with chloroform; 4) fetching the supernate, adding isometric isopropyl alcohol, sodium citrate and sodium chloride and mixing uniformly, and standing at -20 DEG C; 5) after the standing, centrifuging at 4 DEG C, discarding the supernate and leaving the precipitate, adding ethanol prepared by DEPC water, shaking and repeatedly washing and precipitating; and 6) centrifuging at 4 DEG C, discarding the supernate and leaving the precipitate, naturally airing, and adding DEPC water to dissolve RNA to obtain the total RNA solution. The process is simple, the cost is low, the obtained total RNA has high concentration, high purity and high integrity, and further molecular biology study is met.

The invention belongs to the technical field of biology, and particularly relates to an RNA extraction device and method. The device comprises a piston type injector, a plastic centrifugal tube, a centrifugal column arranged in the plastic centrifugal tube and a mixing centrifugal tube, wherein the mixing centrifugal tube comprises a tube body and a tube cover; a through hole is formed in a pipe cover; a silica gel column is arranged in the through hole; a top separation layer is arranged at the upper part of an inner cavity of a pipe body; a middle separation layer is arranged at the middle-lower part of the inner cavity of the pipe body; the inner cavity of the pipe body is divided into an upper cavity, a middle cavity and a lower cavity by the top separation layer and the middle separation layer; grinding balls are pre-placed in the upper cavity; TRIzol is pre-placed in the middle cavity; and chloroform is pre-placed in the lower cavity. According to the invention, an operator doesnot need to directly suck and contact an organic solvent in the RNA extraction process, and toxic volatile matters are not released into an atmospheric environment in the operation process, so the health of experimenters is protected; and meanwhile, mechanical cutting and grinding of blocky tissue samples and difficult-to-split tissue samples are realized through the grinding balls, so RNA is released more quickly and fully.

The invention provides a method for extracting total ribonucleic acid (RNA) of fish testicle tissue. The method includes subjecting the fish testicle tissue to liquid nitrogen quick freezing, trizol lysate treatment, uniform stirring, filtering, centrifugation, digestion by protease K, and extraction by chloroform n-butyl alcohol, precipitation by isopropanol-sodium acetate, performing deoxyribonucleic acid (DNA) enzyme treatment, and performing washing to obtain the total RNA of the fish brain tissue. According to the method for extracting the total RNA of the fish testicle tissue, the influence of a large amount of protein components and connective tissue in the fish testicle tissue can be avoided, and the RNA separation and purification method is stable in purity, good in integrity, high in repeatability, and free of pollution.

The invention discloses an assay kit for testing the relative expression of core-binding factor (CBF) beta / myosin 11 fusion genes, and the kit comprises red blood cell lysis buffer, TRIzol, chloroform, absolute ethyl alcohol, ReverTraAceqPCRRTKit, testing system polymerase chain reaction (PCR) reaction solution, positive control substance and negative control substance, and is characterized in that the testing system PCR reaction solution comprises THUNDERBIRDqPCRMIX, primers of CBF beta / MYH11-F, CBF beta / MYH11-A-R, CBF beta / MYH11-D-R and CBF beta / MYH11-E-R for amplifying a target gene, probe of CBF beta / MYH11-Prob, primers of abl-F and abl-R for amplifying an internal control gene, and probe of abl-Probe. The assay kit can be used for testing the expression level of the CBF beta / myosin 11 fusion genes of a patient suffering from human acute myelognous leukemia (AML), so the test time can be effectively saved, and the test precision is improved.

The invention relates to a method for rapidly and efficiently extracting total RNA of rice tissue. The method is characterized by comprising the following steps of: weighing 100mg of the rice tissue, placing the rice tissue in a porcelain mortar (unsterilized), adding liquid nitrogen into the mortar and quickly grinding the rice tissue, wherein the grinding time is controlled within 1 minute; pouring the ground uniform paste into a 1.5ml EP tube, adding 1ml of Trizol, fully mixing the uniform paste and the Trizol, adding 0.2ml of solution of chloroform into the mixture and fully mixing the mixture for 15 seconds; centrifuging 12,000g of the obtained products for 10 minutes, transferring upper water phase into a new 1.5ml PE tube, adding 0.5ml of isopropyl alcohol and standing the mixture at the temperature of 20 DEG C for 20 minutes; and centrifuging 12,000g of the obtained products for 10 minutes, dumping the supernatant, adding 1ml of newly prepared 75 percent ethanol to wash the precipitate twice, drying the precipitate on a super-clean working table and adding DEPC water to dissolve the precipitate. The extraction method for the total RNA of the rice tissue has the advantages of high speed, high efficiency, completeness and the like.

The invention provides an improved method for extracting total RNA in special vegetable tender leaves. The improved method for extracting the total RNA in the special vegetable tender leaves comprises the following steps: putting an autoclaved and cooled mortar, an autoclave and cooled grinding rod and special vegetable tender leaves into a refrigerator at the temperature of -80 DEG C for more than 4 hours, taking the mortar, the grinding rod and the special vegetable tender leaves out, quickly grinding for about 10s, then sucking right amount of Trizol into the mortar by adopting a pipette, continuously grinding and mixing until jelly is completely dissolved, then sucking the dissolved mixture into a 1.5ml centrifugal tube by adopting a pipette, and then washing, precipitating and carrying out DEPCH2O dissolution, thus the target product, namely the total RNA in special vegetable blades, is obtained. The improved method for extracting the total RNA in the special vegetable tender leaves has the advantages that the problem that RNA is degraded easily (when being placed in the air for a too long time) or a centrifugal tube filled with Trizol mixture bursts (liquid nitrogen is not completely volatilized) as a medicine spoon is used for transferring plant tissue blades smashed by liquid nitrogen into Trizol in the traditional Trizol method for extracting the total RNA in plant tissue blades is solved, the liquid nitrogen is not required to be prepared, and no loss of the ground special vegetable tender leaf powder is produced.

The invention discloses a kit used for detecting relative expression of the AML1-ETO fusion gene. The kit comprises erythrocyte lysate, TRIzol, chloroform, anhydrous ethanol, a ReverTra Ace qPCR RT kit, a detection system PCR reaction solution, a positive control and a negative control, and is characterized in that the detection system PCR reaction solution comprises THUNDERBIRD qPCR MIX, the upstream and downstream primers of AML1-ETO-F and AML1-ETO-R and the probe of AML1-ETO-Probe used for detecting target genes, and the primers of ab1-F and ab1-R and the probe of ab1-Probe used for detecting the internal control gene Ab1. The kit provided in the invention can detect the expression level of the AML1-ETO fusion gene in patients with acute myeloid leukemia (AML) and can effectively save detection time and improve detection precision.

The invention provides a kit for endometriosis diagnosis. The kit comprises 1) an EDTA-K2 anticoagulative tube for collecting peripheral blood, 2) a cryopreservation tube for preloading Trizol, 3) a reagent for extracting total RNA in peripheral blood, 4) a reagent for detecting miRNA-141 and U6 gene expression, and 5) a formula of calculating miRNA-141 scores in blood: miRNA-141 score = miRNA-141Ct value - U6Ct value. The kit has high repeatability and stability and no repeated experiment is required. MiRNA-141 and U6 are utilized to amplify a nucleic acid sequence, the diagnosis kit for determining miRNA score in the blood is constructed, the goal of noninvasive diagnosis of endometriosis can be achieved, the damage to patients caused by laparoscope detection is greatly reduced, and the kit is favorable for early treatment of the endometriosis.

The invention discloses a detection kit for novel coronavirus COVID-19 infection and belongs to the field of in-vitro diagnosis reagents. The kit disclosed by the invention comprises primers and probes for detecting a specific gene ORF1ab of novel coronavirus COVID-19, primers and probes for detecting a positive internal reference gene GAPDH and primers and probes for detecting an exogenous monitoring gene EGFP, and sequences are separately shown in SEQ ID No. 1-9. According to the kit, on the premise that the accuracy and stability of a detection result are guaranteed, the flux of detection is increased. According to the kit, a reagent Trizol is used as a sample preservation solution, safety of living beings, completeness of nucleic acid and load capacity of virus are guaranteed. According to the kit, RNA of the exogenous monitoring gene is introduced during RNA extraction, RNA loss conditions of a whole extraction and detection process can be monitored, the normativeness of a whole operating process is guaranteed, and false negative caused by defects of any step of the operating process is avoided.

The invention relates to a method for detecting expression of microRNA-126-5p in myocardial damage. The method includes following steps: S1, taking out a myocardial tissue specimen collected at minus 80 DEG C, and using Trizol to extract RNA in tissue; S2, reversely transcribing RNA into cDNA: preparing reaction liquid according to a reverse transcription reagent, diluting the reaction liquid, taking and adding diluent into a Real Time PCR system in the next step, and performing quantitative detection; S3, performing RT-QPCR quantitative detection: preparing PCR reaction liquid, and performing Real time PCR, namely S3.1, pre-modifying DNA at 95 DEG C for 30 S, S3.2, performing PCR at 95 DEG C for 5 S and at 60 DEG C for 34 S, and repeating 40 times. The method has the advantage that expression of microRNA-126-5p in myocardial damage can be detected quickly.

The invention belongs to the technical field of environment molecular biology and relates to a method for efficiently extracting microorganism total RNA (Ribonucleic Acid) from anaerobic granular sludge. The method comprises the following steps: fetching activated sludge, centrifuging, discarding supernatant, then adding sterilized glass beads and a stock solution I into a centrifugal tube, putting the centrifugal tube in a vortex mixer for beating after a tube cover of the centrifugal tube is screwed tightly, centrifuging to discharge supernatant, and washing by a phosphate buffer solution; adding a stock solution II and incubating; adding a TRIzol, centrifuging to obtain supernatant after vortex oscillation, adding chloroform, carrying out vortex oscillation, putting in a room temperature, centrifuging to obtain supernatant, adding isopropanol, putting in a temperature of -20 DEG C, centrifuging to discharge supernatanth, adding 75% ethanol to wash precipitates, centrifuging to discharge supernatant, air-drying the precipitates, dissolving nucleic acid precipitates by a stock solution III, adding DNase I (RNase free) and RNase inhibitors and adopting an RNA purifying reagent kit to purify the RNA. The steps for extracting the microorganism total RNA are simplified, and the microorganism total RNA is quickly and efficiently extracted from the anaerobic granular sludge.

The prostate cancerometastasis agent box uses a timely RT-PCR technique, starting from the erythrocytoschisis to separate the surrounding blood individual cell, extracting the overall RNA of each cell through Trizol, reversely transcribing the single cell RNA into CDNA No. 1 chain, achieving timely quantitative RT-PRC inspection through TaqMan detective needle. It improves the sensitivity and astopic feature of PSA and PSMAmRNA inspection, effectively getting over the defects of normal RT-PCR extension, with the precision could detecting one prostate within 107 single cells. This inspection method, design lead, detective needle and clinical sample inspection result provides reliable reference for the development of scheduled quantitative PCR inspection agent box.

The invention discloses a method for preparing a soluble human recombinant MICA protein, which comprises the following steps of: a step 1 of sampling 30ml of in-vitro peripheral venous blood donated by a healthy adult volunteer, diluting the in-vitro peripheral venous blood by physiological saline and separating out an offwhite lymphocyte layer by a lymphocytes separation medium; a step 2 of extracting an RNA (Ribose Nucleic Acid) of processed offwhite lymphocytes by a TRIZOL, carrying out sequence screening on the RNA, planting the RNA into a rhabdovirus genome, carrying out recombination, collection, dialysis and centrifugation to obtain an MICA protein and filtering the MICA protein by a CENTRICON ultrafilter; a step 3 of purifying a target protein; and a step 4 of identifying the MICA target protein. The soluble human recombinant MICA protein prepared by the preparation method is the MICA protein of which a molecular structure domain is closer to the natural state and has the advantages of convenience, rapidness, sensitivity, good repetitiveness and the like. Due to successful application of the soluble human recombinant MICA protein in an in-vitro immunological rejection model, practical support is provided and a solid foundation is laid for in-vivo experiments and clinical application of animals.

The invention discloses a cloning method of a gene NtFLS2 with a function of increasing rutin content of tobacco leaves. The cloning method includes steps: A, using TRIzol for extracting total RNA of tobacco leaves and flowers, and performing reverse transcription to obtain a first strand of cDNA; B, taking the first strand of cDNA as a template, and adopting a primer for PCR (polymerase chain reaction) amplification; C, connecting a target gene segment to a pTOPO carrier to carry out sequencing. The invention further discloses application of the gene NtFLS2 with a function of increasing the rutin content of the tobacco leaves. By cloning of the gene NtFLS2, development and utilization of resources of the gene NtFLS2 lay a foundation for industrial extraction of rutin from the tobacco leaves. According to functional verification of transgenic tobacco plants, the cloned gene NtFLS2 has the function of increasing the rutin content of the tobacco leaves.

The invention discloses a method for extracting RNA from a calcified tissue. The method comprises the following steps: grinding the calcified tissue in liquid nitrogen until the blocky particle diameter is less than 0.5cm, adding Trizol, then adding the liquid nitrogen to continue to grind into powder until the diameter is about 1mm or less than 1mm; then transferring the obtained powder into a RNase-Free tubule, adding the Trizol for cell digestion lysis; centrifuging the tubule, transferring a supernatant to a new tubular; adding chloroform into the supernatant for treatment, then centrifuging, transferring an topmost layer aqueous phase to another new tubular, adding a 75% ethanol solution, mixing gently for nucleic acid precipitation; then transferring to an adsorption column in a kit, standing then centrifuging for RNA to bind to the adsorption column; using a kit solution for washing; and then eluting to obtain the total calcified tissue RNA. The obtained total calcified tissue RNA has the advantages of high purity, high yield, good quality and good integrity.

The invention relates to an animal genetic engineering interferon compound preparation, the production method and the clinical application. The animal genetic engineering interferon compound preparation is prepared by extracting the animal peripheral blood or spleen lymphocytes, culturing, inducing by in vitro inductor, extracting the cell total RNA by Trizol, designing the specific primer, cloning the interferon gene by RT-PCR technology and connecting to the pGEM-T carrier by T-A strategy; the primer is designed again, the both ends are added with different restriction endonucleases digestion sequences, initiation codon ATG and termination codon TAA, leading peptide sequences are removed, the PCR amplification is carried out by taking the recombination T carrier as the template, so as to obtain the expression fragment of the animal interferon gene; the expression fragment is carried out with the rare codon mutation, and gel recovery target fragment is connected on the carrier of pFastBacDual with the same double restriction endonucleases digestion after double restriction endonucleases digestion; the interferon-Alpha of an animal is cloned to multiple cloning sites under the control of Polyhedrin promoter, the interferon-Gamma of the same species animal is cloned to multiple cloning sites under the control of p10 promoter; the constructed carrier of pFastBacDual plus interferon Alpha plus interferon Gamma are transfected to DH10BacE.coli to carry out recombination; the constructed recombinant bacmid is extracted and purified after the blue white screening and the PCR identification, and the transfection of SF9 insect cell is carried out by a liposome method; the expression product is carried out with identification and purification; the animal genetic engineering interferon compound preparation is carried out with anti-virus activity detection and the evaluation of the clinical application effects.

The SCNN1 primer for rapidly diagnosing clear cell nuclear cell carcinoma provided by the invention comprises an internal reference primer pair which is GAPDH, mRNA sequence forward primers and reverse primers of SCNN1A genes with nucleotide sequences shown in SEQ ID NO: 1 and SEQ ID NO: 2, mRNA sequence forward primers and reverse primers of SCNN1B genes with nucleotide sequences shown in SEQ IDNO: 3 and SEQ ID NO: 4, and mRNA sequence forward primers and reverse primers of SCNN1G genes with nucleotide sequences shown in SEQ ID NO: 5 and SEQ ID NO: 6, which take human GAPDH as an internal reference control, and forward primers and reverse primers, having nucleotide sequences shown as SEQ ID NO: 7 and SEQ ID NO: 8. A corresponding kit is also prepared. The kit comprises the following reagents: a tissue lysis solution Trizol, an SYBRGreen Master Mix reaction solution, a reverse transcription reaction solution, a negative control and a positive control. The primer and the kit can eliminate false negative and false positive of clinical sample detection, and are safe, reliable, high in accuracy and suitable for clinical popularization.

The invention provides a method for extracting micro ribonucleic acid (miRNA) from serum. The method comprises the steps that: 500[mu]l of TRIzol is added to 500[mu]l of blood plasma or serum, the mixture is violently concussed for 30s by using a vortex oscillator, and the mixture is then stood under room temperature for 5min; 100[mu]l of isopropanol is added to the mixture, the mixture is well-mixed by tumbling, then the mixture is violently concussed for 30s by using a vortex oscillator until the liquid turns transparent, and the liquid is stood under room temperature for 5min; the liquid is separated for 10min by using a centrifuge under a temperature of 4 DEG C with a speed of 3000rpm, and the supernatant is transferred into a new centrifuge tube; 500[mu]l of chloroform is added to the supernatant in the centrifuge tube, and the liquid is well-mixed by tumbling, then the liquid is separated for 10min by using a centrifuge under a temperature of 4 DEG C with a speed of 13000rpm, and the supernatant is transferred into a new centrifuge tube; ethanol with a volume which is 2.5 times than the supernatant in the centrifuge tube is added to the centrifuge tube, the liquid is well-mixed by tumbling, and the liquid is then stood under room temperature for 10min; the liquid is separated for 10min by using a centrifuge under a temperature of 4 DEG C with a speed of 13000rpm, and then RNA is precipitated; ethanol is removed by absorption, and RNA is dried; the obtained RNA is dissolved with 30[mu]l of DEPC-treated water, and the solution is processed through water-bath for 10min at a temperature of 58 DEG C, such that a target extract is obtained. The method provided by the present invention is simple and applicable. With the method, serum miRNA loss during extraction can be avoided. The method also has advantages of low extraction cost, fast speed and high extraction rate.

The invention provides a method for extracting vibrio parahaemolyticus total RNA, which is characterized by comprising the following steps of: (1) putting 1ml of vibrio parahaemolyticus liquid in a centrifuge tube to be centrifuged; (2) removing supernatant, quickly freezing thallus with liquid nitrogen, grinding into powder, transferring the powder into the centrifuge tube, adding Trizol, shaking for 1min, standing on ice for 3min, and repeatedly shaking for three times; (3) adding chloroform / isoamylol, and centrifuging; (4) transferring 500ul of supernatant into the centrifuge tube, adding 500ul of isopropanol, standing to fully precipitate and centrifuging; (5) carefully removing supernatant, washing with 70 percent of ethanol, and centrifuging; and (6) carefully absorbing supernatant as far as possible, inversely standing on a ultra-clean table to dry alcohol, adding DEPC water for dissolving, and storing under the temperature of 80 DEG C below zero. Since adopting the technical scheme, the invention has the advantages and the beneficial effects that the method can extract total RNA with high concentration, high purity and good integrity in the short three hours, and the used reagent is simple and has low cost.

The invention discloses a kit for detecting relevant expression quantity of an AML-EVI1 (acute myelocytic leukemia-ecotropic virus integration site-1) fusion gene. The kit comprises red blood cell lysis buffer, TRIzol, chloroform, absolute ethyl alcohol, RevertraAceqPCRRTKit, detecting system PCR (polymerase chain reaction) liquid, a positive reference substance and a negative reference substance. The kit is characterized in that the detecting system PCR reaction liquid comprises THUNDERBIRDqPCRMIX; a target gene is detected via upstream and downstream primers AML1-EVI1-F and AML1-EVI1-R and a probe AML1-EVI1-Probe; and an internal control gene Abl is detected via primers abl-F and abl-R and a probe abl-Probe. The kit disclosed by the invention can detect the expression level of the AML1-EVI1 fusion engine in a patient body subjected to CML (chronicmyelognousleukemia), thus, the detecting time can be effectively salved, and the detecting precision can be increased.