87results about How to "Short amplification time" patented technology

The invention discloses a H3 subtype flu quick-detecting type classifying method based on an RT-LAMP technique. The method comprises the following steps: firstly, analyzing the HA gene sequence of the H3 subtype flu virus in a flu virus gene group sequence database, designing highly special and conserved RT-LAMP special primers and six primers corresponding to eight areas of target genes, wherein four primers are FIP, BIP, F3 and B3, and the other two primers are loop primers LP1 and LP2; secondly, treating a sample to be detected in advance, and extracting the total RNA of the sample to be used as a reaction template; thirdly, configuring an RT-LAMP reaction system; fourthly, mixing the RNA template into the RT-LAMP reaction system, arranging the RT-LAMP reaction system in a water bath, and carrying out RT-LAM enlargement at constant temperature; and fifthly, identifying the enlarged reaction product. The result shows that the reaction product contains H3 subtype flu virus when the reaction product is positive; and the reaction product is H3 subtype flu virus when the reaction product is negative.

The invention provides a method and system for individual recognition and paternity identification of an unknown sample. The method includes: extracting the DNA of the unknown sample; acquiring the typing result of 17 loci contained by the DNA, i.e. 14 autosome STR loci, 2 Y-chromosome loci, and the sex determination locus Amelogenin, with the STR loci being D6S1043, D21S11, D7S820, CSF1PO, D2S1338, D3S1358, D13S317, D8S1179, D16S539, Penta E, D5S818, vWA, D18S51 and FGA, the Y-STR locus being DYS448, and the Y-chromosome insertion/deletion site being M134; and carrying out individual recognition and paternity identification according to the genotypes of the 17 loci of the unknown sample. The invention also provides the system for individual recognition and paternity identification of the unknown sample. The scheme involved in the invention can realize simultaneous detection of autosome STR loci and Y-chromosome loci, and also can shorten the detection time, thus improving the efficiency of individual recognition.

The invention discloses a culture medium for expanding human mesenchymal stem cells, which comprises a basic medium and an additive added to the medium, and the additive includes the following components at final concentrations: linoleic acid 2‑10 μL/mL, human Source AB type serum 5‑15 μL/mL, human transferrin 5‑15 μg/mL, etc. Using the culture medium to expand human mesenchymal stem cells includes the following steps: (1) isolating human mesenchymal stem cells from umbilical cord or placenta, or isolating mononuclear cells from bone marrow blood, umbilical cord blood or placental blood, and placing them in the culture medium (2) Place the cells separated and purified in step (1) into fresh medium for subculture. The invention adopts the culture medium of specific components and the expansion method, which can effectively speed up the cell expansion speed, shorten the expansion time, maintain the uniform shape of the cells after passaging, and reduce their differentiation, thereby ensuring the safety of human mesenchymal stem cells sex.

The invention relates to a method for breeding lean-type Chinese Huai pigs in a multi-gene pyramiding manner based on growth traits thereof. The method comprises the following operating steps: 1, carrying out DNA (deoxyribonucleic acid) extraction on the genome of the pig; 2, designing primers, more particularly, designing the PCR (polymerase chain reaction) amplification primers according to the gene sequences of insulin-like growth factor-I (IGF-I), pituitary specific transcription factor-I (PIT-I), liver X receptor alpha (LXR alpha) and melanocortin-4 receptor (MC4R); 3, carrying out the PCR; 4, carrying out restriction fragment length polymorphism (RFLP); and 5, carrying out the correlation analysis on the polymorphism and growth traits of genes. According to the analysis, the method can determine that the single growth rate of the pyramided gene with the gene type thereof being AADDGGFF is the highest one; the method can prevent the genes of other offsprings from being isolated after the selective reservation for breeding, thus achieving the optimal effect of the growth traits of the offsprings on the four gene types; and the method can stabilize the inheritance, particularly lead the genes of the growth traits to become homozygous within one generation, thus accelerating the cultivation of the lean-type Chinese Huai pigs.

The invention discloses an inhibition method for induced differentiation of hair follicle stem cells into vascular endothelial cells, comprising: (1), isolating rat hair follicle stem cell; (2), culturing the rat hair follicle stem cells; (3), purifying the rat hair follicle stem cells; (4), inhibiting the induced differentiation of the rat hair follicle stem cells into vascular endothelial cells. In the inhibition method provided herein, vascular endothelial cell growth factor 165 is used for the first time as an inducing factor so that the rat hair follicle stem cells efficiently and directionally differentiate into vascular endothelial cells; the formation and growth of vascular endothelial cells generated by induced differentiation of the hair follicle stem cells can be adjusted, an effective healing of a wound is promoted; it is also possible to provide seed cell sources for solving the problems in vascularization of tissue engineering skin, cell-transplant therapy of ischemic disease and the like.

The invention relates to a primer composition, kit and method for detecting the novel coronavirus. The primer composition includes a first primer pair and a second primer pair which are used for detecting the N gene of the novel coronavirus, and also includes a third primer pair and a fourth primer pair which are used for detecting the ORFlab gene of the novel coronavirus. The method comprises the steps: using the primer composition, adopting the genomic DNA of a to-be-tested sample as a template, performing a constant-temperature amplification reaction under the action of thymine DNA glycosylase, MLV reverse transcriptase and Bst DNA polymerase, and performing quantitative analysis on the test results under the action of fluorescent dye so as to detect the novel coronavirus with high specificity and high sensitivity. The four primers are used, bases of the primers are specially modified, and the thymine DNA glycosylase and Bst DNA polymerase are added to the reaction, so that exponential amplification of target sequences of the N gene and the ORFlab gene are realized under the condition of constant temperature, and the sensitivity and specificity of detection are improved greatly.

The invention discloses a culture medium capable of promoting the division growth of mesenchymal stem cells, which comprises a serum-free basic culture medium and an additive added on the basis of theserum-free basic culture medium, wherein the additive comprises a hibiscus mutabilis extract, seaweed polysaccharide, astragaloside IV, human serum albumin, transferrin, glutamine, platelet-derived factor, epidermal growth factor, fibroblast growth factor, human insulin growth factor and vitamin A; the hibiscus mutabilis extract has antioxidant and cell activating activity, and can effectively promote the growth of cell metabolic by compounding seaweed polysaccharide and astragaloside IV; human serum albumin, transferrin, glutamine and vitamin A provide essential nutrient substances for the growth of stem cells; platelet-derived factor, epidermal growth factor, fibroblast growth factor, human insulin growth factor and other cytokines together promote the rapid growth and proliferation ofstem cells; the culture medium not only can improve the growth activity of mesenchymal stem cells, shorten the culture time, promote the expression of cell growth factors, but also can maintain the stem cell activity of the differentiation potential of mesenchymal stem cells; stem cells are not differentiated in daily culture, thereby providing convenience for scientific research.

The invention belongs to the technical field of edible fungi cultivation and specifically relates to a cultivation method for oyster mushrooms. The cultivation method comprises the following steps: S1, strain activation; S2, strain amplification; S3,inoculation; S4, hairy fungus cultivation; S5, mushroom producing management; and S6 harvesting. A culture medium is prepared from reasonable raw materials, and a nutrient solution is sprayed after former-stubble mushrooms are harvested, so that the yield of the oyster mushrooms is remarkably increased.

The invention relates to the technical field of biological gene detection, and aims to provide an ovarian premature senility related gene whole exon amplification and detection method. The method comprises the following steps: extracting genome DNA from a detection material; using totally 120 primers shown in SEQ ID NO:1-120, carrying out an amplification test on all exons of 9 genes, namely FMR1,FOXL2, FSHR, POF1B, INHA, NOBOX, GDF9, BMP15 and FIGLA, wherein totally 61 PCR reactions in the amplification test and the negative control are carried out synchronously, and the Tm value range of the 120 primers is 57-62 DEG C; detecting an amplification product by adopting agarose electrophoresis and a gel imaging system; and analyzing the sequencing result of the amplification product by usingan automatic sequencer. All exons of 9 genes related to premature ovarian failure are synchronously amplified through optimized PCR amplification specificity, and a detection raw material is providedfor downstream detection. The method is simple and convenient to operate, short in amplification time, high in specificity and good in repeatability. The detected positive rate is greatly improved, and the application prospect is wide.

The invention relates to a gene chip, a primer set and a kit for single-tube detection of non-deletion alpha-thalassemia and beta-thalassemia, and belongs to a molecular diagnosis technology. According to the present invention, based on the direct multiplex PCR and reverse dot blot combined detection principle, the corresponding amplification primers and the corresponding probes are designed according to the mutation or deletion sites of each genotype, the primer is labeled with biotin, the probe is labeled with amino, a gene chip is used as substrate, the probe is immobilized on the DNA chip,the PCR product amplified by the specific primer is hybridized with the probe, and the diagnosis of thalassemia is performed by interpreting a signal coloring box.

The invention relates to a detection method for sensitively and rapidly detecting staphylococcus aureus enterotoxin B, comprising the following steps: heating H1 and H2 with concentration of 5Mum at 95 DEG C for denaturation for 5min, heating an SEB aptamer at 85-95 DEG C for denaturation for 5-10min, then taking 6.7-33.3muL of SEB aptamer with concentration of 1muM, adding 100muL of SEB, mixing and then adding 80-100muL of hybrid buffer solution, and reacting for 30min at 37 DEG C; adding 6.7-33.3muL of 1muM H0, and reacting for 30min at 37 DEG C; adding 10-30muL of mixed reaction system of 100muL of H1 with concentration of 5muM and 100muL of H2 with concentration of 5muM; reacting for 45min at 37 DEG C constant temperature, and measuring the fluorescence value; compared with a conventional detection technology based on nucleic acid aptamer, the detection sensitivity is reduced by 2 orders of magnitudes.

The invention discloses a CDA primer group and a kit for detecting African swine fever virus (ASFV), and an application of the CDA primer group and the kit. The CDA primer group is a primer group for amplifying a conserved region fragment of the African swine fever virus, namely ASFV-MF-3/ASFV-MR-3, wherein the nucleotide sequence of the ASFV-MF-3 is as shown in SEQ ID NO. 5, and the nucleotide sequence of the ASFV-MR-3 is as shown in SEQ ID NO. 6. The primer group provided by the invention is high in sensitivity and strong in specificity, and the kit prepared from the primer group can quickly and accurately detect whether a sample to be detected contains the African swine fever virus or not. In addition, the visual kit provided by the invention can provide great convenience for on-site detection in places such as farms, customs, ports and community vegetable markets with low professional degrees.

The invention provides a primer composition for detecting clostridium difficile. The primer composition is designed based on a conserved sequence of a toxin gene tcdB of the clostridium difficile. Theinvention further provides a reagent kit for detecting the clostridium difficile, and the reagent kit comprises the primer composition, enzymes for identifying and excising an unconventional DNA basein a strand in double-strand DNA, and DNA polymerase having the function of strand displacement. The invention further provides a method for detecting the clostridium difficile. According to the method provided by the invention, target sequences can be correctly detected, the specificity is good, and the detection sensitivity and the detection efficiency are greatly improved.

The invention discloses a serum-free medium for expanding human mesenchymal stem cells, which is based on the medium for culturing stem cells, adding linoleic acid, human AB type serum, and platelet-derived growth factors. The invention also discloses a culture method for human mesenchymal stem cells. The serum-free medium of the present invention uses multiple cytokines in combination reasonably, which not only improves the purification effect of mesenchymal stem cells, but also shows that the morphology of human mesenchymal stem cells is still uniform even after 30 passages, thus ensuring The safety of human mesenchymal stem cells improves the expansion speed of cells, reduces the expansion time and reduces the cost.

The invention relates to a mycoplasma pneumoniae (MP) rapid detection primer group and a kit. The mycoplasma pneumoniae rapid detection primer group comprises an upper stream primer, a lower stream primer and a probe which detect a mycoplasma pneumoniae gene group P1 gene sequence. The kit comprises a lysate solution, a reconstitution fluid, an EMA reaction tube containing primer probes, a positive quality control product and a negative quality control product. The primer group can be specifically combined with the mycoplasma pneumoniae P1 gene, and cooperate with EMA (Enzymes Mediated Amplification, a multienzyme mediated nucleic acid amplification technology, namely a normal temperature nucleic acid amplification technology) to prepare the mycoplasma pneumoniae detection kit, and the mycoplasma pneumoniae rapid detection primer group and kit have the advantages of easy to operate, short in consumed time, high in sensitivity and specificity and the like.