59 results about "Guanidine isothiocyanate" patented technology

The invention relates to the technical field of nucleic acid extraction and purification, in particular to a DNA and RNA co-extraction reaction reagent and an extraction method, the reaction reagent comprises a nucleic acid protective agent RP, a sample lysis binding solution LB, a protein washing solution WB1, a nucleic acid rinsing solution WB2 and a nucleic acid eluent EB; the nucleic acid protective agent RP comprises DTT, mercaptoethanol, TCEP and sodium hydroxide; the sample cracking binding solution LB comprises Tris-HCl, benzenesulfonic acid, sodium citrate, guanidine hydrochloride, guanidine isothiocyanate, guanidine thiocyanate, urea, triton 100, SDS, potassium chloride, sodium chloride, bromhexine, N-acetylcysteine, ambroxol hydrochloride, isopropanol and absolute ethyl alcohol; the protein washing liquid WB1 comprises Tris-HCl, benzenesulfonic acid, sodium citrate, guanidine hydrochloride, guanidine isothiocyanate, guanidine thiocyanate, urea, triton 100, sodium chloride, isopropanol and absolute ethyl alcohol; the nucleic acid rinsing liquid WB2 is prepared from Tris-HCl, sodium chloride, isopropanol and absolute ethyl alcohol; and the nucleic acid eluent EB is nuclease-free water. The reaction reagent can be used for rapidly and sufficiently enriching and purifying DNA and RNA of a sample at the same time.

The invention relates to the technical field of nucleic acid detection, in particular to a lysis buffer solution, a kit and an extraction method for extracting free DNA of peripheral blood. The lysis buffer solution for extracting the free DNA of the peripheral blood comprises a first lysis solution and a second lysis solution, the first lysis solution takes guanidine hydrochloride as a main chaotropic agent and at least one of ammonium thiocyanate and guanidine isothiocyanate as an auxiliary chaotropic agent, and is compounded with a metal ion chelating agent, trehalose, dithiothreitol, digestive enzyme, sodium dodecyl sarcosinate, borate and ascorbic acid; the second lysis solution takes guanidine hydrochloride as a main chaotropic agent and urea as an auxiliary chaotropic agent, and is compounded with a metal ion chelating agent, trehalose, dithiothreitol, digestive enzyme, sodium dodecyl sarcosinate, borate, ascorbic acid and polyethylene glycol. The buffer solution of the lysate does not contain SDS (sodium dodecyl sulfate), so that the residue of SDS in an extracted product can be avoided, and the amount of extracted free DNA (deoxyribonucleic acid) is more.

The invention provides a pretreatment reagent and a method for nucleic acid extraction of an excrement sample. The pretreatment reagent is a solution comprising TritonX-100, PEG6000, SDS, guanidine isothiocyanate, KAl (SO4) 2 and ammonium acetate. According to the reagent provided by the invention, the scheme of the excrement pretreatment reagent capable of realizing one-step pyrolysis and inhibitor sedimentation can be realized, the operation process and time of manual uncovering and sample transfer are greatly shortened, pyrolysis, flocculation and flocculation termination are all completed in the same tube, only one centrifugation step is needed, and the clinical operability is greatly improved. Meanwhile, the scheme has more excellent performance than a commercial excrement pretreatment reagent.

The invention relates to the technical field of virus extraction kits, in particular to a large-volume sample virus nucleic acid rapid extraction kit and method.The large-volume sample virus nucleic acid rapid extraction kit comprises a protease K solution, magnetic beads, a lysis solution, a rinsing solution 1, a rinsing solution 2 and an eluent; the magnetic beads are hydroxyl magnetic beads; the lysate is prepared from guanidine isothiocyanate, Tris alkali, Tris hydrochloric acid, triton X-100, isopropyl alcohol and polyoxyethylene ether; the rinsing liquid 1 is composed of a guanidine hydrochloride solution and isopropanol; the rinsing liquid 2 is composed of absolute ethyl alcohol and purified water; the kit provided by the invention can easily obtain viral nucleic acid from a large-volume sample, is simple to operate, can extract a plurality of samples at one time, realizes rapid diagnosis, and makes up the vacancy of a kit for automatically extracting viral nucleic acid from a large-volume sample at high flux in the current market.

The invention provides a kit for extracting plant genome DNA based on magnetic beads and application. The kit is composed of a lysis solution, a binding solution, a washing solution I, a washing solution II, an eluent and magnetic beads. The cracking combination solution comprises at least one of lauryl sodium sulfate, cetyltrimethylammonium bromide, guanidine hydrochloride, guanidine isothiocyanate, citric acid, trisodium citrate, Tween 20, polyvinylpyrrolidone, sodium chloride, ethylenediamine tetraacetic acid and Tris-HCl; the cracking combination solution comprises at least one of sodium dodecyl sulfate, cetyltrimethylammonium bromide, guanidine hydrochloride, guanidine isothiocyanate, citric acid, trisodium citrate, Tween 20, polyvinylpyrrolidone, sodium chloride, ethylenediamine tetraacetic acid and Tris-HCl; the binding solution comprises at least one of isopropanol, ethanol, guanidine hydrochloride and polyethylene glycol; the washing liquid I is at least one of guanidine hydrochloride, guanidine isothiocyanate, absolute ethyl alcohol, ethylenediaminetetraacetic acid, sodium chloride and Tris-HCl, the washing liquid II is an ethanol aqueous solution, the eluent is a Tris-HCl and ethylenediaminetetraacetic acid aqueous solution, and the magnetic beads are hydroxyl magnetic beads or carboxyl magnetic beads.

The invention discloses an injector-based nucleic acid purification device, an extraction method and a viral nucleic acid lysate. The purification device comprises an injector assembly and a filter membrane assembly, wherein the injector assembly comprises an injector tube barrel and a push rod matched with the injector tube barrel; the filter membrane assembly is connected to the outlet end of the injector tube barrel, and the filter membrane assembly is configured to be of a detachable structure. The nucleic acid purification device provided by the invention perfects the processes of nucleic acid release, nucleic acid washing and elution, can remove components which interfere with subsequent PCR (Polymerase Chain Reaction), fluorescent quantitative PCR and constant-temperature detection in the nucleic acid release process, and can improve the detection efficiency; the viral nucleic acid lysate is suitable for common clinical samples such as nasal swabs, anus swabs, serum and ascites, and is suitable for nucleic acid extraction and purification of DNA and RNA viruses by optimizing the proportion of guanidine isothiocyanate, polyvinylpyrrolidone (PVP-40) and lauryl sodium sulfate (SDS).

The invention discloses a nucleic acid extraction reagent combination. The nucleic acid extraction reagent combination is prepared from the following components: MagZol 3BD, Buffer BCP2, Buffer RW1 and Buffer RW2, the MagZol 3BD is mainly prepared from guanidine isothiocyanate, phenol and ultrapure water; the Buffer BCP2 is mainly prepared from 1-bromo-3-chloropropane, and the buffer BCP2 is prepared from 1-bromo-3-chloropropane; the Buffer RW1 is mainly prepared from guanidine isothiocyanate, ethanol and ultrapure water; and the Buffer RW2 is a 10 * STE buffer solution. The invention also discloses a nucleic acid extraction kit containing the nucleic acid extraction reagent combination, and application of the nucleic acid extraction kit in screening and diagnosis of prostatic cancer. The nucleic acid extraction reagent combines a one-step RNA extraction technology and a column RNA purification technology, can be used for blood nucleic acid extraction and prostatic cancer diagnosis, detects the expression levels of TIMP1, TMEM176B and ITGB5 genes related to prostatic cancer in blood through blood total RNA extraction and a multiple fluorescent PCR technology, and diagnoses prostatic cancer through a three-gene risk prediction model. The kit has very high specificity and sensitivity for diagnosis of the prostatic cancer, and can be used for effective screening and diagnosis of the prostatic cancer.

This invention relates to the field of fecal genomics kit technology, and particularly to a safe, rapid, and high-throughput kit for extracting fecal genomes and its usage method. The kit includes a sample preservation solution, magnetic beads, a lysis buffer, a washing buffer 1, a washing buffer 2, and an elution buffer. The sample preservation solution is composed of guanidine isothiocyanate, sodium citrate, Triton X-100, Tween 20, propidium iodide, and purified water. The magnetic beads are hydroxyl magnetic beads. The lysis buffer is composed of guanidine hydrochloride, Tris base, Tris hydrochloric acid, Triton X-100, isopropanol, and purified water. Washing buffer 1 is composed of guanidine hydrochloride solution and isopropanol. Washing buffer 2 is composed of anhydrous ethanol and purified water. This kit can easily extract the genome from fecal samples, is simple to operate, safe and non-toxic, and can extract multiple samples at once, enabling rapid diagnosis and filling the gap in the current market for automated high-throughput kits for extracting genomes from fecal samples.

The invention discloses a kit and a method for extracting helicobacter pylori nucleic acid in an excrement sample and application. The kit comprises sample treatment liquid: guanidine isothiocyanate and lithium dodecyl sulfate; the lysis solution is prepared from magnetic beads, guanidine isothiocyanate, lauramidopropyl hydroxysulfobetaine, potassium chloride, trihydroxymethyl aminomethane and isopropanol; the washing liquid I is prepared from guanidine hydrochloride, Tween 20 and tris (hydroxymethyl) aminomethane; a washing liquid II: betaine, guaiacol and ethanol; the washing liquid III is ethanol; the eluent is tris (hydroxymethyl) aminomethane. When the kit is used for extracting the helicobacter pylori nucleic acid in the excrement sample, impurities can be fully digested, adsorption of the impurities and the magnetic beads is reduced, cross-linking of the impurities and the nucleic acid is reduced, effective adsorption of the nucleic acid and the magnetic beads is increased, and the yield and quality of the helicobacter pylori nucleic acid in the excrement sample are improved.

The invention belongs to the technical field of biology, and provides a mycobacterium tuberculosis detection method and a kit, the detection method comprises the following steps: adding a to-be-detected sample containing mycobacterium tuberculosis into a lysis working solution, carrying out a heat preservation reaction, centrifuging, and taking a supernatant, the cracking working solution contains a biotinylated metal complexing agent and biotinylated nanoparticles, and guanidine isothiocyanate and hydrophobic alkyl groups are connected to the biotinylated nanoparticles; adding a magnetic bead suspension into the supernatant, stirring at room temperature for reaction, standing, taking an upper solution, adding the upper solution into a purification column, purifying and washing to obtain a detection sample, and the surface of a magnetic bead contains streptavidin; and adding a detection sample into an amplification reagent for fluorescent PCR amplification, and analyzing a PCR amplification product by using a fluorescence melting curve to finish detection. The method can effectively reduce impurities contained in the detection sample and improve the detection accuracy.

The present invention is applicable to the field of biotechnology and provides a method for extracting Helicobacter pylori DNA from feces, which comprises the following steps: collecting a fresh fecal sample of a patient infected with Helicobacter pylori and placing it in a centrifuge tube; adding a cell lysate to the centrifuge tube and performing homogenization. The cell lysate includes: Tris-HCl, ethylenediaminetetraacetic acid, guanidine isothiocyanate, Triton X-100, isopropanol, sodium dodecyl sulfate, sodium chloride, proteinase K, modified polyethylene glycol, and modified betaine. By adding modified polyethylene glycol to the cell lysate, the capture of free DNA is enhanced, promoting further concentration and enrichment of DNA and improving the subsequent binding efficiency of DNA to magnetic beads. By adding modified betaine, the effects of removing impurities such as proteins and esters are improved, reducing the impurities adsorbed by magnetic beads in the subsequent steps. Modified polyethylene glycol and modified betaine have a synergistic effect and can improve the purity of the extracted DNA.

The invention discloses a plasma free DNA extraction reagent, a kit and an extraction method, the extraction reagent comprises protease K, a lysis solution, a binding solution, a washing solution 1, a washing solution 2, a washing solution 3 and an eluent, the extraction reagents contain more common and less-dosage common reagents such as Tris-HCl (pH 8.0), EDTA.2Na. 2H2O, NaCl, glycerol, SDS, isopropanol, ethanol, Tween-20 and the like. The method has the advantages that the problems that the reagent is easy to separate out and needs to be heated and redissolved for use during storage can be solved without traditional high-concentration guanidine salts such as guanidine hydrochloride and guanidinium isothiocyanate, the stability and the operation convenience of the extraction reagent are increased, a better extraction effect is achieved, and the eluent can effectively protect the extracted cfDNA from being degraded.

The present application relates to the technical field of nucleic acid extraction and purification, and particularly relates to a DNA and RNA co-extraction reaction reagent and an extraction method, the reaction reagent comprising a nucleic acid protective agent RP, a sample lysis and binding liquid LB, a protein washing liquid WB1, a nucleic acid rinsing liquid WB2, and a nucleic acid elution liquid EB; the nucleic acid protective agent RP comprising DTT, mercaptoethanol, TCEP, and sodium hydroxide; the sample lysis and binding liquid LB comprising Tris-HCl, benzenesulfonic acid, sodium citrate, guanidine hydrochloride, guanidine isothiocyanate, guanidine thiocyanate, urea, triton X-100, SDS, potassium chloride, sodium chloride, bromhexine, N-acetylcysteine, ambroxol hydrochloride, isopropyl alcohol, and anhydrous ethanol; the protein washing liquid WB1 comprising Tris-HCl, benzenesulfonic acid, sodium citrate, guanidine hydrochloride, guanidine isothiocyanate, guanidine thiocyanate, urea, triton X-100, sodium chloride, isopropyl alcohol, and anhydrous ethanol; the nucleic acid rinsing liquid WB2 comprising Tris-HCl, sodium chloride, isopropyl alcohol, and anhydrous ethanol; and the nucleic acid elution liquid EB being nuclease-free water; the reagent can rapidly and fully enrich and purify DNA and RNA of a sample simultaneously.

The invention provides a magnetic bead method liquid sample pathogenic microorganism nucleic acid extraction kit, which is characterized by comprising protease K, a nucleic acid protection solution, a lysis strengthening solution, a lysis binding solution, magnetic beads MXVB, a washing solution 1, a washing solution 2 and an eluent, wherein the cracking combination liquid is prepared from tris (hydroxymethyl) aminomethane, guanidine isothiocyanate, ammonium thiocyanate, tween-80, lauryl sodium sulfate, sodium chloride and isopropanol; the nucleic acid protection liquid comprises tris (hydroxymethyl) aminomethane hydrochloride and formamide; the cracking reinforcing liquid is lauryl sodium sulfate. According to the kit, DNA of bacteria, fungi and viruses can be rapidly extracted and purified, the extraction efficiency of pathogenic microorganism nucleic acid is improved, good stability is achieved, the nucleic acid quality is guaranteed, and development of a pathogenic microorganism detection technology is facilitated.

The present application relates to the technical fields of nucleic acid extraction kit, and particularly relates to a high-purity whole blood genomic nucleic acid extraction kit, which comprises a protease K solution, magnetic beads, a lysis solution, rinse solution 1, rinse solution 2, rinse solution 3, rinse solution 4 and an elution solution; the lysis solution is composed of guanidine isothiocyanate, guanidine hydrochloride, Tris alkali, EDTA and isopropyl alcohol; the magnetic beads are hydroxyl magnetic beads; the rinse solution 1 is composed of sodium chloride, isopropyl alcohol and purified water; the rinse solution 2 is composed of Triton X-100, NP-40, sodium chloride and purified water; the rinse solution 3 is composed of anhydrous ethanol, guanidine isothiocyanate and purified water; the rinse solution 4 is composed of anhydrous ethanol and purified water; and the elution solution is purified water; the kit can easily obtain genomic nucleic acid from a whole blood sample, is simple to operate, can extract multiple samples at a time, realizes rapid diagnosis, is safe and non-toxic, and fills the gap of the automatic high-throughput genomic nucleic acid extraction kit for whole blood samples in the current market.

The invention relates to a reagent combination or kit for rapidly extracting free nucleic acid from a trace plasma sample and an extraction method, and belongs to the technical field of biological diagnosis. The reagent combination or the kit comprises the following components: a sample treatment solution, a rinsing solution, magnetic beads and an eluent, the sample treatment liquid is prepared from the following main components: guanidine isothiocyanate, urea, Tris-HCl, EDTA (Ethylene Diamine Tetraacetic Acid), NaCl, Tween-20, Triton-100, ammonium sulfate and isopropanol; the main components of the rinsing liquid are Tris-HCl and absolute ethyl alcohol. The main component of the eluent is water. The method for extracting the free nucleic acid in the trace plasma sample is simple and convenient to operate, high in efficiency, less in reagent combination, more economical and capable of realizing automation, the content of inhibitors in the obtained nucleic acid is low, and downstream experimental analysis is facilitated.

The invention relates to the technical field of biology, in particular to a nucleic acid extraction reagent composition and application thereof. The invention provides a reagent combination simultaneously adaptive to common nucleic acid extraction and electrowetting platform adaptive nucleic acid extraction, according to the reagent combination provided by the invention, high-concentration guanidinium isothiocyanate is added into a lysis solution, heating is performed in a lysis step to assist lysis, lysis and nucleic acid adsorption are synchronously performed at the same time, and the extraction time is shortened; a surfactant with proper concentration is added into the lysis solution, the washing solution and the eluent, so that the reagent combination can be better driven on an electrowetting platform, the driving force is improved, and the uniform mixing effect is improved; meanwhile, protease K is added into the washing liquid, the common step of washing protein with high-concentration salt in a magnetic bead extraction method is omitted, a good washing effect can be achieved only through one-step washing, the extraction time is further shortened, meanwhile, residual protease K in the washing liquid is inactivated through high-temperature elution, follow-up amplification is not affected, and the method is suitable for application and popularization.

The invention discloses a method for efficiently and rapidly extracting nucleic acid based on a paramagnetic particle method, and belongs to the technical field of molecular biological detection. A lysis solution used in the method is prepared from the following components: 1 to 4 M of guanidine isothiocyanate, 0.5 to 1.5 percent of Triton X-100, 0.2 to 1 percent of Tween 20, 10 to 50 mM of EDTA (Ethylene Diamine Tetraacetic Acid), 10 to 50 mM of DTT (Diethylene Terephthalate), 4 to 8 percent of PEG (Polyethylene Glycol) 8000 and 1 to 3 M of NaCl. According to the method, ethanol with gradient concentration is used for washing. According to the method disclosed by the invention, the optimized lysis solution is combined with gradient concentration ethanol for washing, so that rapid lysis of cells and efficient extraction of nucleic acid can be realized within 10 minutes; according to the method, volatile organic reagents such as chloroform and Trizol do not need to be used, so that the personal safety of operators is guaranteed; the method is simple in process operation, low in professional requirement on experimental operators and suitable for various complex samples such as plasma and serum, the technical application range is expanded due to wide compatibility, and diversified research requirements are met.

The invention provides a method for releasing gamma-retroviral vector nucleic acid in PG13 cell culture supernatant and a releasing agent thereof. The method comprises the following steps: step 1, carrying out enzyme digestion on a cell supernatant sample containing a gamma-retroviral vector produced by packaging cell PG13 culture; 2, a nucleic acid releasing agent is added into the sample subjected to enzyme digestion in the step 1, nucleic acid releasing is conducted, and the nucleic acid releasing agent is composed of guanidine isothiocyanate, absolute ethyl alcohol, isopropyl alcohol and enzyme-free water. The method disclosed by the invention is suitable for a sample nucleic acid release method in which host DNA needs to be removed and virus vector nucleic acid is RNA, and has no interference effect on subsequent quantitative detection. According to the method, the nucleic acid release effect of first enzyme digestion and then release is obviously better than that of an existing method of first nucleic acid release and then enzyme digestion.

The invention discloses a stool sample preservation solution and its application, wherein the preservation solution contains the following ingredients: guanidine isothiocyanate, Tween 80, PEG-8000, NaCl and anhydrous ethanol. The stool sample preservation solution prepared by the invention adopts a special formula, which is different from the traditional stool sample preservation solution. The traditional stool sample preservation solution can usually only be preserved for 7-10 days at room temperature (15°C-30°C). The preservation solution of the invention can quickly release sample nucleic acid at room temperature (37±2)°C, and can stably preserve nucleic acid for at least 30 days, which greatly extends the storage period of stool samples, is more conducive to the transportation of samples and related detection in the later stage, and has the advantages of low cost and convenient use.

The invention relates to a kit for extracting DNA from FFPE tissue. The kit comprises a dewaxing solution, a lysis solution and a denaturation solution, the dewaxing liquid contains mineral oil, dimethyl silicone oil and butyl laurate; the lysis solution contains sucrose, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride, ethylenediamine tetraacetic acid, polysorbate-20 and water; the denaturation liquid is prepared from guanidine isothiocyanate, trihydroxymethyl aminomethane salt, sodium chloride, ethyl phenyl polyethylene glycol, triton X-100, lauryl sodium sulfate and water. According to the kit, the use of a traditional toxic organic reagent dimethylbenzene is avoided, and dewaxing liquid, lysate, denaturation liquid and an extraction method are improved, so that the nucleic acid fragmentation is reduced, the nucleic acid yield is increased, the genome DNA can be safely, quickly and efficiently extracted from an FFPE sample, the yield and integrity of the DNA are improved, and the kit is suitable for downstream DNA sequencing analysis.

The invention belongs to the technical field of molecular detection, and discloses a nucleic acid hybridization capture immunofluorescence detection kit as well as a use method and application thereof. In the kit provided by the invention, guanidine hydrochloride, guanidine isothiocyanate, triton X-100, NaCl, protease K and lauryl sodium sulfate are compounded according to a specific concentration to obtain the nucleic acid release liquid, so that pathogen nucleic acid in a clinical sample can be fully dissociated, and the nucleic acid release liquid can cooperate with a probe aiming at a specific target pathogen; the solution system can be used for optimizing the recognition and combination process of DNA and RNA, the specific recognition and combination capacity between the probe and specific pathogen nucleotide is improved, and under the condition that a common nucleic acid stabilizer, namely Tween, is not contained, the solution system can further strengthen differential structural epitopes existing in a DNA / RNA complex compared with DNA and RNA, so that the specificity of the DNA / RNA complex is improved, and the specificity of the DNA / RNA complex is improved. The kit can be better recognized and combined by an anti-DNA / RNA complex antibody, and has excellent detection specificity, sensitivity and precision.

This invention relates to the field of biological sample preservation technology, specifically to a room-temperature preservation solution for bacterial RNA from sputum samples of tuberculosis patients. Each 100 mL of the RNA preservation solution comprises the following ingredients: 2-5 g tris(hydroxymethyl)aminomethane, 0.5-1.3 g mucin digester, 20-30 g guanidine isothiocyanate, 0.1-0.3 g hexadecylpyridine chloride, 0.5-1.2 g ethylhexene dicysteine, 20-40 mL fixative, 1-5 g stabilizer, 0.1-0.5 g inorganic salt, 0.3-0.6 g surfactant, and 0.1-0.5 g preservative. The preservation solution provided by this invention can effectively liquefy sputum samples at room temperature, meeting the requirements for long-distance sample transportation. The RNA quality of sputum samples treated with the preservation solution of this invention is suitable for various subsequent molecular detection methods, possessing significant application value and promising prospects for widespread application.

The application relates to a sample processing method for cervical exfoliative cell PCR detection, which adopts a sample processing reagent, including a processing liquid, a protection liquid, a conversion liquid, a first purification liquid, a second purification liquid, a third purification liquid and a collection liquid. The processing liquid contains 0.1-0.5N sodium hydroxide and 4-8mol / L urea; the protection liquid contains 1-8mM guanidine isothiocyanate, 0.1-5% SDS, 5-150mM Tris-Hcl and 5-100mM EDTA; and the conversion liquid contains 100-500mM sodium bisulfite, 50-250mM ammonium sulfite, 2-15M ammonium bisulfite, 0.1-3M tetrahydrofurfuryl alcohol, 1-5mM TEPA and 0.1-2M magnesium sulfite. The sample is lysed and protected by the processing liquid and the protection liquid, direct conversion and detection are facilitated, processing time and steps are saved, DNA loss is reduced, cost is saved, the accuracy and reliability of detection results are ensured.

The invention discloses a free miRNA (micro Ribonucleic Acid) extraction lysis buffer and an extraction method, the miRNA extraction lysis buffer comprises the following components: 0.1 mol / L to 0.5 mol / L of Tris-HCl, 3 mol / L to 6 mol / L of guanidine isothiocyanate, 0.15 mol / L to 0.30 mol / L of potassium chloride and 2.00 mol / L to 5.30 mol / L of N, N-dimethylacetamide. The N, N-dimethylacetamide is added into the miRNA extraction lysis buffer based on guanidinium isothiocyanate, so that the yield of miRNA can be improved, the sensitivity of a clinical sample miRNA detection reagent is improved, the miRNA extraction stability is remarkably improved, miRNA expression quantity deviation caused by a nucleic acid extraction link is reduced, and finally the quantitative accuracy of sample miRNA is improved.

The invention relates to the technical field of methylation detection, and particularly discloses a plasma sample-based rapid methylation pretreatment kit, which comprises a lysis solution, magnetic beads, a rinsing solution and a conversion solution, the lysis solution is prepared from Tris-HCl, EDTA (Ethylene Diamine Tetraacetic Acid), NaCl, SDS (Sodium Dodecyl Sulfate), guanidine isothiocyanate, sodium acetate, PVP (Polyvinyl Pyrrolidone), PEG200 ( The magnetic beads are polydisperse magnetic microspheres with the particle size of 100-400 nm; the magnetic bead rinsing liquid is an ethanol solution; according to the method, the formula of the extraction reagent is improved, the protease K is successfully removed on the premise that the extraction efficiency is not changed, the specific conversion reagent is adopted, the large volume of the specific conversion reagent serves as the extraction eluent, and conversion can be directly carried out after elution. The innovative design effectively reduces the loss of nucleic acid and obviously improves the detection sensitivity. Meanwhile, the extraction process and the conversion process are integrated, so that the operation steps are greatly simplified, the operation complexity is reduced, and a more efficient and convenient sample pretreatment solution is provided for methylation detection.

The invention provides a rapid and efficient shell separation method for shellfish larvae, which relates to the technical field of shell separation for shellfish larvae, and comprises the following steps: S1, obtaining scallop larvae, and washing with 3x PBS solution; S2, dissociating larvae with lysis solution, and separating shells and soft tissues, wherein the lysis solution contains the following components: guanidine isothiocyanate, sodium lauroyl sarcosinate, and sodium citrate; S3, collecting and washing larval shells; S4, detecting the degree of separation of larval shells. By adopting the rapid and efficient shell separation method for shellfish larvae of the invention, the separation time is greatly shortened, the damage to the structure and components inside the shell is reduced, and the soft tissue of the shell after separation is less polluted. In addition, the larval shell separated by the method can extract complete DNA, and the high-throughput sequencing library is qualified. The invention effectively solves the technical problem of shell separation for shellfish larvae, and provides a basis for related research on genomics, transcriptomics, proteomics, metabolomics, etc. of larval shells.

The invention provides gastric juice sample preserving fluid as well as a preparation method and application thereof, and belongs to the technical field of biology. The gastric juice sample preserving fluid takes water as a solvent and comprises the following components: 0.5 to 4.5 M of guanidine isothiocyanate, 4.5 to 55 mM of disodium ethylene diamine tetraacetate, 0.5 to 22 mM of tris (2-carboxyethyl) phosphine and a tris (hydroxymethyl) aminomethane buffer solution. The preservation solution can improve the detection effect of the gastric juice sample, can effectively protect the gastric juice sample for 5 days at room temperature, has the effect equivalent to that of-20 DEG C cryopreservation, and provides more flexibility and convenience for transportation and preservation of the sample.

This invention provides a nucleic acid extraction reagent, kit, and method. The nucleic acid extraction reagent includes a pretreatment solution, proteinase K, lysis binding buffer, magnetic beads, washing buffer 1, washing buffer 2, washing buffer 3, and elution buffer, all stored separately but used in combination. The pretreatment solution has a pH of 7.80-8.20 and is composed of 0.1%-1% sodium dodecyl sulfate, Tris-HCl solution, and EDTA-Na2 solution by mass. The lysis binding buffer has a pH of 7.80-8.20 and is composed of 3M-4M guanidine isothiocyanate, 11%-15% polyethylene glycol 6000 by mass, 5%-10% Triton X-100 by mass, Tris-HCl solution, and EDTA-Na2 solution. This invention achieves "high concentration, high purity, and high efficiency" nucleic acid extraction through innovative extraction reagent components (non-toxic lysis system), reagent kit design (high-throughput pre-packaging), method optimization (automated coupling), and application expansion (adaptation to frozen samples), providing a reliable template for molecular diagnostics.

The invention relates to the technical field of biology, in particular to a lysis solution for rapidly extracting bacterial nucleic acid and a rapid nucleic acid extraction method based on the bacterial lysis solution and magnetic bead separation and enrichment. The lysate is prepared from 50m of Mtris (pH = 7.5), 20mM of EDTA (Ethylene Diamine Tetraacetic Acid), 0.7 M of NaCl, 1.5 M of guanidine isothiocyanate, 1M of guanidine hydrochloride, 0.2 percent of SDS (Sodium Dodecyl Sulfate), 1.5 percent of Tween-20, 1.5 percent of Triton X-100 and 1 percent of PEPPG F68. The extraction method comprises the steps of sample pretreatment and cracking, magnetic bead combination, magnetic bead washing, magnetic bead redissolution and magnetic bead separation, and can be used for treating Gram-positive bacterium and Gram-negative bacterium samples. Silicon hydroxyl magnetic beads with specific particle sizes are adopted, a 2mM NaOH solution is used for washing, and a Tris-HCl (pH = 7.9) solution is used for redissolving. The lysate can be used for preparing a reagent for rapidly extracting bacterial nucleic acid, the extraction method can be applied to bacterial detection or molecular biology experiments, the nucleic acid extraction time is effectively shortened to be within 10 minutes, and an efficient and reliable technical means is provided for related research and detection work.