31results about How to "The experimental steps are simple" patented technology

The invention discloses a high-effective construction method for a T carrier based on a PCR method. The invention is that a pair of oligodeoxynucleotide sections which contain specific restriction enzyme sites are introduced into an initiation carrier (such as pUC18); the restriction enzyme site is characterized in that the corresponding enzyme (such as EcoRV) can generate blunt end, and the baseof fro and post tangency point are respectively T and A; the initiation carrier is linearized under the function of the enzyme which is then used as a PCR template; designed primers produce two single chain through single-primed PCR or asymmetric PCR amplification, the two single-chains are annealed so as to form the T carrier. The method for preparing the T carrier is simple and fast; the produced T carrier can effectively clone PCR products containing tail A, and the non-reconstitution background is nearly zero.

The invention provides a high-efficiency recombined adenovirus containing an ndrg2 gene and the pharmic purpose thereof. The recombined adenovirus contains the ndrg2 gene, a Gateway system and a Virapower TM Adenoviral Expression system are utilized in the construction process, and the adenovirus is packed by a recombination method in vitro. After being cloned to an introduction carrier, a target gene can be efficiently and quickly cloned to other carriers of a receptor by specific recombination sites on the carriers and recombinase without depending on a restriction enzyme. The package of the adenovirus in vitro simplifies the experiment steps, shortens the operation time, and has high multiplication efficiency, high transgene efficiency and high virus yield efficiency as well as the transduction efficiency as high as 100 percent. The invention solves the technical problem of recombination and package of the ndrg2 gene in vitro as well as probes and determines the lethal effect of the high-efficiency recombined adenovirus to brain glioma cells and cells of lung cancer, liver cancer, gastric cancer and colon cancer. The high-efficiency recombined adenovirus can be used as a medicament for treating brain glioma, long cancer, liver cancer, gastric cancer and colon cancer.

The invention discloses a basalt fiber surface treatment device and method.The treatment device comprises a drying oven, a treatment liquid atomization mechanism, a control signal generator and a basalt fiber fixing assembly, the treatment liquid atomization mechanism is installed at the top of the drying oven, and the basalt fiber fixing assembly is installed on one side plate of the drying oven; the control signal generator is installed on the edge of the other side plate of the drying oven, the working end of the basalt fiber fixing assembly corresponds to the atomization spraying end of the treating fluid atomization mechanism, and the treating fluid atomization mechanism and the basalt fiber fixing assembly are electrically connected with the control signal generator. By using the device, the basalt fibers can be quickly cleaned, etched, treated by a coupling agent and dried. The system is compact in layout, automatic in adjustment, convenient and fast, automatic control is achieved according to a set program, experiment steps are greatly simplified, processing time is saved, and processing efficiency and result success rate are improved.

The invention discloses a synthesis method of a camphenyl-based Schiff base compound. The specific synthesis steps are as follows: taking camphenal as a raw material, adding aromatic amine compounds to a reaction device in an organic solvent, and molar amounts of camphenal and aromatic amine The ratio is 1:1-1.1 for feeding, stirring and reflux for 2-3 hours, the product after the reaction is purified with a solvent, and then dried to obtain camphenyl Schiff base compounds. The preparation method of the present invention uses amphene aldehyde as a raw material, reacts with aromatic amine compounds to obtain Schiff base compounds, introduces amphene groups into the Schiff base, and finally obtains amphene group-containing aromatic amine Schiff bases, and the experimental steps are simple and convenient. The reaction condition is mild, the time is short, the yield is high, the reaction is safe and environmentally friendly, the post-treatment is convenient, and the secondary pollution is small.

The invention provides a method for preparing 2-amino-5-(N-phenothiazinyl)methylene-1,3,4-thiadiazole. The2-amino-5-(N-phenothiazinyl)methylene-1,3,4-thiadiazole is obtained by the steps of adding N-carboxyl-methylene-phenothiazine, thiosemicarbazide and concentrated hydrochloric acid into a reactor, then refluxing to obtain a reaction mixture, cooling the reaction mixture to a room temperature, regulating a pH value of the reaction mixture to 8-9 by using a sodium hydroxide aqueous solution, then cooling the reaction mixture until solid particles are precipitated, suction pumping the solid particles to obtain a filter cake, washing the filter cake by ice water and carrying out recrystallization. The method is simple in experimental step and post-treatment, high in yield and small in secondary pollution, is an economic and environment-friendly industrial preparation method, and fits to need of industrial production.

The invention discloses a method for synthesizing alpha-amino ketone from ketone and imine, and belongs to the technical field of chemical organic synthesis. The method comprises the following steps: adding imine into an organic solvent, sufficiently dissolving so as to obtain an imine solution, further respectively adding ketone, a catalyst tetrabutyl ammonium iodide and an oxidant tert-butyl hydroperoxide into the solution, sealing up a tube opening, heating to be 130 DEG C, stirring for 3 hours, cooling down the system to be the room temperature after the reaction is ended, extracting by using dichloromethane and removing the solvent of on organic phase so as to obtain alpha-amino ketone. By adopting the method, defects in the conventional method for synthesizing alpha-amino ketone by electrophilic amination and nucleophilic amination are overcome, direct amination of alpha-position of a simple ketone compound such as acetone is achieved, a C-N bond is generated through oxidative coupling reaction between a N-H bond and a C-H bond at one step, and the alpha-position of a ketone substrate does not need to be prefunctionalized, so that the experiment steps are simple, the condition is gentle, the reagents are cheap, and the range of the substrate is wide; the method is applicable to industrial production.

The invention discloses a method for measuring the temperature of a hydrocarbon flame based on a colored CCD camera. According to the method, the temperature measurement range of a measured object is not limited by a blackbody furnace calibration experiment, and a blackbody furnace is not used. The method comprises the following steps: (1) photographing the hydrocarbon flame, namely setting parameters of the colored CCD camera, and photographing the flame; (2) acquiring a relation curve between the temperature and a specific value of original strengths of two base colors, namely acquiring a relation curve between the temperature and a specific value of the original strengths recorded by channels R, G and B; (3) acquiring R, G and B true color images, namely outputting RAW format images from the colored CCD camera, and finally converting the RAW format images into the R, G and B true color images; (4) acquiring a specific value of the two base colors, namely acquiring the original strengths of the three channels R, G and B from the R, G and B true color images, and calculating the specific value of the original strengths of the optional two base colors; and (5) acquiring the temperature, namely acquiring the temperature of the hydrocarbon flame according to the obtained relation curve between the temperature and the specific value of the original strengths of the two base colors.

The invention discloses a Mannich alkali compounds containing phenothiazine group and 1,3,4-thiadiazole group as well as a preparation method and an application thereof. The preparation method comprises the following steps of: stirring and reacting a phenothiazine, formaldehyde solution and 1,3,4-thiadiazole derivatives used as raw materials in the presence of ethyl alcohol used as a solvent at a room temperature; then steaming out the solvent in the obtained reaction solution, and recrystallizing the remainder, namely, a crude product, in a reactor, so as to obtain Mannich alkalis containing phenothiazine group and 1,3,4-thiadiazole group. The preparation method disclosed by the invention is simple in operation steps, convenient in aftertreatment, and high in yield; moreover, the compounds can be applied to preparation for anti-gram bacteria medicines.

The invention relates to a method for preparing modified HEK293T cells, the modified HEK293T cells, a method for preparing drug-loaded exosomes, and drug-loaded exosomes. The method for preparing and transforming HEK293T cells includes the following steps: S1: Plasmid construction: using the pcDNA3.1 expression vector as a plasmid backbone, respectively constructing three plasmids: pcDNA3.1-STEAP3, pcDNA3.1-SDC4, and pcDNA3.1-NadB fragments ; S2: The three plasmids in S1 were transfected into HEK293T cells according to the ratio of 1-3:1-3:1-3, and the stable cell lines were screened to obtain the transformed HEK293T cell lines. By adopting the above technical scheme, due to the synergistic effect of these three plasmids, HEK293T cells that secrete a large amount of exosomes can be transformed, thereby further increasing the amount of exosomes secreted and providing more options for clinical trials.

The invention relates to a method for high-efficiency detection of rhodamine B in food and a rapid detection kit. The detection method is mainly divided into three steps: 1, extracting rhodamine B in food; 2, purifying and enriching the rhodamine B in food; and 3, detecting through a machine. The corresponding kit consists of an extracting agent 1, an extracting agent 2 and a solid extraction column 3. When the rhodamine B in food is detected by employing the method, the detection limit can reach 0.5ppb, and the external standard recovery rate is over 85 percent. The food is subjected to pretreatment by employing the method and the key reagents such as the extracting agent 1 and the extracting agent 2, so that the rhodamine B in food is completely extracted, the toxicity of the used reagent is obviously reduced, the required time for detecting the rhodamine B in food is greatly shortened, the detection precision is improved, and the method has the excellent characteristics of high convenience, rapidness, environmental friendliness and high applicability and implementability, and a toxic agent is avoided.