70results about How to "Shortened culture" patented technology

The invention relates to a method for high-efficiency fermentation production of L-glutamic acid. The method can solve the problems of lower acid production, low glucose-acid conversion rate, and the like of the prior production process. The method comprises the following steps: a biotin auxotroph strain is taken as a production strain, according to a certain proportion, a plurality of nutritional components are prepared into a culture medium with rich and balanced nutrients as the fermentation culture medium, the dissolved oxygen is controlled on the appropriate level through the regulation of the stirring speed of a fermentation tank and the wind amount, the pH is controlled by feeding ammonia water, and the residual sugar is controlled on the lower level by feeding glucose solution with a certain concentration, and the fermentation is stopped till 32h. The method can shorten the whole fermentation cycle and greatly improve the yield (more than 150g/l) and the conversion rate (more than 60 percent) of the L-glutamic acid under the situation of not increasing any additional equipment and manpower investment, the whole process has simple operation and lower production cost, thereby being very applicable to industrial production.

The invention relates to the field of microorganisms, and in particular discloses a test tube screening method of an isaria cicadae strain. The method comprises the following steps: collecting wild cordyceps cicadae or conidium thereof, and separating and purifying through a panel culture medium to obtain isaria cicadae single colony; picking the isaria cicadae single colony from the panel culture medium in the former step to a slope culture medium, and culturing to obtain conidium; collecting the conidium obtained from the former step, preparing spore solution, and inoculating to a test tube equipped with a test tube culture medium made of wheat; culturing and observing sporocarp, spore or hypha production condition. The test tube culture is adopted by the test tube screening method of the isaria cicadae strain, the occupied culture area is small, the operation is convenient, the culture period is short, the seed culture process in a shake flaks is not needed, the labor intensity is greatly reduced, and the energy consumption in the screening process is reduced; the wheat culture medium has good isaria cicadae strain selective culture effect.

The invention relates to a stem cell repairing material as well as a preparation method and application thereof. The stem cell repairing material comprises a stem cell and a cell carrier, wherein the stem cell is an autologous adipose-derived mesenchymal stem cell (ADSC) with a concentration of 105-108/ml, and the cell carrier is autoserum, normal saline (0.9%) or a glucose injection (5%). In the invention, adopted seed cells are prepared through separating and purifying autologous adipose tissues of patients, thereby avoiding the ethical disputes and the immunological rejection. The material drawing of the adipose tissues can be performed by using an instrument suction method, which is simple in operation and can bring less traumas and pains to the patients. The cell carrier used in the invention is the autoserum, normal saline (0.9%) or glucose injection (5%), therefore, the cell carrier mixed with mesenchymal stem cells and other ingredients can be injected into the patents by virtue of syringes. By using the stem cell repairing material provided by the invention, the operation process is simple, no scar is left, and the wounds and pains for the patients are avoided, therefore, the stem cell repairing material can be easily accepted by the patients.

The invention belongs to the technical field of biology, in particular to a haematococcus pluvialis cell growth promoting agent and a use method thereof. Concretely, rhodofix dissolved by ethanol and 3-seradix dissolved by hydrochloric acid are diluted into distilled water to obtain the promoting agent, wherein each liter of the solution has the rhodofix content between 0.5 and 3.0 ppm, and the 3-seradix content between 1.0 and 5.0 ppm. The obtained promoting agent is added into a culture medium of the haematococcus pluvialis cells. The invention uses the characteristic that the plant growth regulating substances have the features of promoting the cell division, the cell propagation and the growth in a certain concentration range. Through the analysis on the effect of the cell growth speed of different haematococcus pluvialis strain cells under different plant growth regulating agents and concentration matching processing, the grow promoting agent with the improved broad-spectrum effect on the haematococcus pluvialis cell growth and the proper concentration of each ingredient can be obtained through integration and optimization. The haematococcus pluvialis cell growth promoting agent of the invention has simple use method, and has the obvious promoting effect on the haematococcus pluvialis cell growth speed and the astaxanthin accumulation.

The invention relates to the technology of plant tissue culture, and in particular relates to a tissue culture method for inducing populus yunnanensis dode tetraploid in combination with colchicine. The method taking petiole of populus yunnanensis dode as a tissue culture material in the process of tissue culture is characterized by comprising the following steps of: adding colchicine into a populus yunnanensis dode petiole differential medium to induce populus yunnanensis dode to generate polyploidy, wherein the treatment time is 20-30 days; carrying out differentiation culture again in a populus yunnanensis dode petiole differential medium in which colchicine is not added after the treatment; cutting the part which is obvious in variation on the differentiated cluster buds; repeatedly cultivating on the differential medium for 5-6 times, so that the populus yunnanensis dode petiole differential medium is stable in polyploidization variation; and performing rooting on the clump seedlings which are stably varied, so that the tissue culture seedlings of the tetraploid populus yunnanensis dode can be obtained. The method provided by the invention is the tissue culture method for inducing the populus yunnanensis dode tetraploid in combination with the colchicine, which is easy and simple to operate and can effectively reduce the chimera.

The invention discloses a method for identifying the resistance of tobacco to root rot caused by Fusarium tabacinum. The method comprises 1, culture of Fusarium tabacinum, 2, preparation of a Fusariumtabacinum liquid, 3, cultivation of tobacco seedlings, 4, pre-treatment on the tobacco seedlings, 5, inoculation with the Fusarium tabacinum, and 6, investigation to detect the resistance of the specie to the root rot caused by Fusarium tabacinum according to the disease conditions. The identification method can directly identify tobacco seedlings. Compared with the method for inoculation of a pot or a field with bacteria, the method is free of a complex process for pot culture after transplant, reduces work load, utilizes floating plate vaccination to inoculate with hundreds or thousands ofplants, enlarges the inoculation amount, greatly reduces identification time through direct inoculation in the seedling period, breaks the season limitation of the resistance of tobacco to root rot caused by Fusarium tabacinum and realizes identification in any seasons.

The invention discloses a microbiological method based harmlessness treatment technology for agricultural wastes generated by grape cultivation. The method comprises steps of seed culture, liquid state fermentation and solid state fermentation. A Trichoderma strain is Trichoderma viride or Trichoderma harzianum. The invention not only provides a novel approach for harmlessness treatment and resource utilization of molasses alcohol wastes from sugarcane processing and a large amount of agricultural wastes generated by grape cultivation, besides, a solid Trichoderma agent obtained from the treatment can be applied to vineyards to increase biomass of Trichoderma for biological control in the environment, improve microecology and soil of the vineyard, inhibit pathogenic microorganisms for grapes in the microecology of the vineyard and reduce initiation factors for microbial diseases of grapes, thereby reducing application amount of pesticide, and enhancing fruit quality and security of the grapes.

The invention provides a microfluidic perfusion culture device, and the device comprises a fluid control unit, a culture chamber unit and a substrate liquid path unit; the fluid control unit connectsa culture medium into the substrate liquid path unit through a pump body, and the substrate liquid path unit controls liquid path selection through on-off of an electromagnetic valve, so the culture medium flows into a culture chamber; and circulating perfusion culture of the culture chamber is realized. According to the system, waste of manpower and resources can be reduced, shunting control overliquid, long-time high-flux perfusion culture of microorganisms or cells and real-time observation and detection are automatically achieved, culture and detection of the microorganisms or the cells are easy to operate, space is saved, and the system can be used in the ground or aerospace microgravity environment.

The invention relates to a preparation method of artificial seeds of submerged plants (hydrilla). The method comprises the following steps: 1. washing the taken hydrilla seedling with water to remove plankton on the surface of the hydrilla seedling; 2. cutting off the roots and leaves of all the plants, picking up the eugenic plants and washing the eugenic plants with distilled water; 3. cutting off the lateral buds and the terminal buds with internodes and putting the lateral buds and the terminal buds into gel liquid which is formed by mixing of sodium alginate, hoagland's nutrient solution and NAA (nicotinic acid amide); 4. dropping the gel liquid containing hydrilla buds into CaCl2 by a dropper to perform curing reaction, taking out gel liquid after the curing reaction and rinsing with sterile distilled water, and terminating the reaction to obtain the artificial hydrilla seeds. According to the method, the step of plant tissue culture is eliminated, and the method is not required to be implemented under an aseptic condition, the preparation process is greatly reduced, the cost is greatly reduced, the period is shortened, and the survival rate is high.

The invention provides a method for direct extraction of rice blast pathogen DNA from rice leaf infected by rice blast or neck blast single scab. The method of the invention can directly extract rice blast pathogen DNA from rice leaf or neck infected by rice blast, the extraction method is simple, fast and low-cost, and the extracted DNA is applicable to PCR based analysis on rice blast pathogen nontoxic gene detection and colony genetic diversity.

The invention provides a new method for fast selecting engineering bacteria of leaven of efficient expression antimicrobial peptides. The method comprises the following steps: 1. constructing plasmidsfor syncretizing antimicrobial peptide genes and chaperone protein genes; 2. carrying out enzyme restriction for linearizing the plasmids; 3. transforming the plasmids to pichia competent thalli through electroporation, utilizing auxotroph for selecting, and cultivating the plasmids for 36 to 48 hours at the temperature of 28 DEG C; 4. directly obtaining transformants from a prescreening plate, i.e. obtaining the engineering bacteria of the leaven of the efficient expression antimicrobial peptides primarily, and then the engineering bacteria of the leaven can be obtained from large number oftransformants through reselection. The invention has simple experiment process and easy operation, the entire screening process can be shortened by 5 to 6 days, and the expenses of labor, materials and finical resources are greatly reduced. The invention is suitable for the engineering bacteria expressed in the leaven for proteins of the antimicrobial peptides, enzyme, antibodies, polypeptide andthe like which is not easy to or can not adopt a functional approach for selection.

The invention discloses a preparation method of skin fibroblasts, and belongs to the technical field of biological cells. The preparation method of the skin fibroblasts comprises the following steps of: S01, specimen treatment: treating a skin specimen, and removing adipose tissues and connective tissues; S02, separation: treating the treated specimen with neutral protease, and removing epidermis to obtain dermis tissues; S03, digestion: digesting the dermis tissues by using pancreatin to obtain tissue fluid; S04, centrifugation: centrifuging the tissue fluid, and removing supernatant to obtain a cell suspension; S05, culture: transferring the cell suspension into a culture dish, adding a culture solution, and culturing; and S06, passage: culturing until the fusion degree is 75-85%, and performing passage culture. The preparation method disclosed by the invention has the advantages of simple steps, low possibility of pollution, a high cell survival rate, a good purification effect and small cell loss.

The invention provides a rapid propagation method for a clustered shoot through induction of the aerial root of Anthurium andraeanum Lind. The method comprises the following steps: 1) selection of an explant; 2) induction culture of reproduction block mass; 3) induction culture of a clustered shoot; 4) propagation of the clustered shoot; and 5) acclimatization and transplant, wherein an obtained seedling is transplanted into a pot in two months so as to obtain a tissue culture seedling bred by using the method of propagation of the clustered shoot through induction of the aerial root of Anthurium andraeanum Lind. According to the invention, the method overcomes the disadvantages of a low propagation coefficient and a complex rooting culture method in induction of a clustered shoot through reproduction block mass in the prior art; hormones consisting of TDZ and NAA are utilized to induce regeneration of the clustered shoot from the aerial root of Anthurium andraeanum Lind, so a high propagation coefficient is realized, rooting and acclimatization procedures are simplified, the reproduction block mass of the aerial root has superior differentiation capability and anti-aging capability compared to calluses induced by different materials, the purposes of a fast propagation speed and conservation of production cost are achieved, and the method is applicable to large scale seedling growing in a factory.

The invention discloses an N<15> stable isotope labeling method for biological protein quantifying and tracing. The method comprises the steps that yeast is utilized for converting an N<15> inorganic salt into biological protein, wherein when the culture passage number of the yeast reaches 6 or above, 98% or above of protein extracted from the yeast is N<15> protein; the high-purity N<15> protein extracted from the yeast is converted into N<15> yeast peptone, the N<15> yeast peptone is utilized for replacing conventional N<14> yeast peptone to be used for culturing multiple organisms, and specific N<15> isotope labeling is provided for biological protein labeling quantifying and tracing. The method can be applied to more organism varieties and is easy and convenient to operate, low in experimental cost and short in test cycle, and people can conduct proteome analysis on a large number of samples in an efficient and low-cost mode conveniently.

The invention discloses a biosynthesis method of ademetionine. The method comprises the following steps: subjecting ademetionine engineering bacteria to fermentation culture so as to obtain fermentation broth, filtering the fermentation broth, drying the wet bacteria for 2 to 3 hours at a temperature of 60 to 65 DEG C, then activating the dried bacteria, adding adenine, glucose, phosphate, sulfate, methionine and water into the obtained activated bacteria to form a reaction system, and carrying out conversion reactions at a temperature of 33 to 37 DEG C, when the conversion reactions finish, the reaction liquid is a mixed solution containing ademetionine. An SAM engineering bacterium is constructed, and can add methionine in a reaction liquid of an ATP synthesis reaction through adenine, so the steps of SAM synthesis enzyme culture, SAM synthesis enzyme extraction, and SAM synthesis enzyme addition during the reaction process are saved, and the production is more convenient. Moreover, the raw materials such as adenine can be bought from the market at a low price in a large amount. The biosynthesis method of ademetionine can efficiently synthesize SAM in a large scale.

The invention discloses an RNA probe for detecting novel coronavirus 2019-nCOV as well as a preparation method and application thereof, and belongs to the technical field of nucleic acid hybridization. In order to improve the accuracy and specificity of novel coronavirus detection, the invention provides the RNA probe for detecting novel coronavirus 2019-nCOV, and the nucleotide sequence of the RNA probe is shown as SEQ ID No.1. The preparation process comprises the following steps: primer designing, construction of recombinant plasmids, PCR amplification, product purifying, and in-vitro transcription. When the prepared probe is applied to detection of 2019-nCOV, 2019-nCOV virus nucleic acid can be detected at a single cell level, and the detection sensitivity is improved.

The invention provides a method for direct extraction of pathogenic fungus and bacteria DNA from leaf scabs of gramineous crops such as pathogenetic rice and corn. The method of the invention can directly extract pathogen DNA from leaf and neck of rice and corn leaf scabs infected by rice blast, the extraction method is simple, fast and low-cost, and the extracted DNA is applicable to PCR based analysis on pathogenic organism molecular detection and diagnosis, pathogen nontoxic gene detection and colony genetic diversity.