51 results about "Rapid dna" patented technology

Dermatophytes which belong to one of the three genera Epidermophyton, Trichophyton and Microsporum are the main cause of fungal infections of skin, hair and nails. Traditional diagnostic procedures consist of microscopy and culture, but due to the slow growth rate of dermatophytes typically two to four weeks are needed before a final diagnosis is obtained. The present invention is a rapid DNA extraction method extracting nucleic acids from fungi (e.g. dermatophytes and other pathogenic fungi) which can be performed from directly on hair, nail or skin specimens from humans, from naturally or experimentally infected animals or from cultured fungal colonies for the use in PCR amplification and detection assays. The present invention also includes specific primer sets for detection of any dermatophyte and for species specific detection of Trichophyton rubrum and Epidermophyton floccosum by PCR and a kit for diagnosing fungal infections.

The present invention concerns microfluidic systems with printed surface structured electronically controllable wettability switches for efficient manipulation of small amounts of fluids. The high performance microfluidic systems of the invention can be used in many applications, e.g. in rapid DNA separation and sizing, cell manipulation, cell sorting and molecule detection.

The invention discloses a rapid DNA extraction kit of a medicinal material, belongs to the field of the molecular biology, and relates to the traditional Chinese medicine and traditional Chinese medicinal material identification field. The kit comprises a solution A, a solution B, a reagent C and a user manual; and the solution A is composed of a solute triton-100 and a solvent water, the concentration of the solute is 1%, and the final concentration of the solute in the solution A 1%. The solution B is composed of 1M of a solute Tris-HCl and the solvent water, the concentration of the Tris-HCl in the solution B is 1M, the pH value of the solution B is 8.0, the concentration of the solute is 0.1M, and the final solution of the solute in the solution B is 0.1M. The reagent C is composed of NaOH and polyvinylpyrrolidone (PVP, M=40), the final concentration of NaOH in the solution A is 0.5M, and the final concentration of the PVP in the solution A is 1%. The kit is a medicinal material genome DNA extraction product developed for rapidly discriminating the medicinal material, and can be used for rapidly extracting plant and animal medicinal material genome DNA. The kit has the advantages of few operation steps, fast extraction speed, low price, no need of a large device, large-scale high-flux extraction and the like, and is very suitable for rapid detection and identification of the medicinal material DNA.

The invention relates to the field of molecular biology and particularly discloses a specific primer pair for amplifying a gene D8S1179. The specific primer pair comprises D8S1179-FP:5'-TTTGTATTTCATGTGTACATTCGTAATTC-3' and D8S1179-RP:5'-ACCTATCCTGTAGATTATTTTCACTGATTG-3'. The invention further provides an STR typing method for a loca D8S1179 based on next generation sequencing and particularly provides a primer combination for being connected with a proper adaptor. The STR typing method is applied to the STR typing of the loca D8S1179. According to the primer pair capable of amplifying the gene D8S1179, after concentrations of different sample products subjected to PCR amplification, the equivalent mixing is carried out, and a genome library is constructed according to a BIOO RAPID DNA kit; the library is input into a computer by virtue of a next generation sequencer, data is subjected to bioinformatics analysis, and the typing results such as times of repetition of NDA molecules with same length, SNP of the loca and the difference among side wings of the loca of D8S1179 can be found.

The invention relates to an application of a plant leaf grinding device for high-throughput rapid DNA extraction, and belongs to the technical field of biology. The grinding device comprises: a pedestal, a handle, and a plurality of grinding heads and connecting rods, wherein the pedestal is cuboid, is made of a thermal insulation material, and is a device for fixing the connecting rods and the handle; the upper part of the pedestal is connected to the handle; the lower part of the pedestal is connected to the connecting rods; and the lower parts of the connecting rods are connected to the grinding heads. The 96 grinding heads are connected to the base of the pedestal through the 96 connecting rods in an arrangement manner of 8 rows and 12 lines, wherein the transverse distance between two adjacent connecting rods is 0.37-0.9 cm and the longitudinal space distance between two adjacent connecting rods is 0.37-0.9 cm to match a 200 muL 96-hole PCR board and a 96-hole PCR device. For the application of the plant leaf grinding device for high-throughput rapid DNA extraction, whole sample crushing on the 96-hole PCR board can be completed in one step; a requirement of rapid DNA extraction and direct PCR experiment is met; and grinding time is greatly saved and labor requirement is lowered.

The invention discloses a simple and rapid DNA extraction method in wine, which comprises the following steps: A. 5g Silica is added with 5ml aqueous suspension for Silica precipitation to be evenly mixed after being cleaned by pure water and is set at the temperature of 4 DEG C for standby; B. wine with best adsorption conditions is then produced; C. the DNA of the wine is adsorbed and recycled: (1) Silica suspension is added to the wine mixture; (2) conservation is done for 1-2 hours under the temperature of 4DEG C, with constant shaking and oscillation; (3) centrifugation is done for 5 min under the temperature of 4DEG C with the amount of 5000g; (4) supernatant is removed and the Silica deposit is washed for 1-2 times by using washing fluid containing 0.1M Tris-HCl, pH 8.0, 1 percent of CTAB and 1M NaCl; (5) the centrifugation is done for 5 min with the amount of 5000g to collect the Silica deposit; (6) the Silica deposit is added with the pure water with the equivalent amount as the deposit and then well shaked and done with DNA heat elution after 5-10 min thermal insulation at the temperature of 65 DEG C; (7) centrifugation (10000g, 5min) is done to collect the supernatant containing DNA; (8) the supernatant in step (7) is extracted by chloroform with the same volume and the centrifugation (10000g, 3min) is done to collect top phase containing DNA; (9) the DNA in the supernatant is recycled and concentrated by using isopropanol precipitation; (10) the recycled wine DNA is then dissolved in 50 Mu lota TE (10mM Tris-HCl, pH 8.0) buffering agent.

The invention discloses a novel rapid DNA separation and sequencing method for a mitochondrial genome and a kit. A linear chromosome sequence is degraded by Exonuclease III or Plasmid-Safe ATP-Dependent DNase, and then mitochondrial genome DNA is enriched for high-throughput sequencing. The mitochondrial DNA sequencing method is applied to detection of the mitochondrial genome and particularly detection of the mitochondrial genome without a consensus sequence.

The present invention relates to detection of citrus diseases and particularly relates to a rapid DNA extraction method of citrus leaves, and an application thereof in a high-throughput molecular detection of citrus yellow shoot diseases. The rapid DNA extraction method comprises the following steps: petioles and leaf midribs of the citrus leaves from citrus plants to be tested are cut and crushed, 40 [mu]L of lysate is added into each 30 mg of a sample, vortexing vibrating is conducted for 10-15 s, water bath at 95 DEG C is conducted for 15 s, then 0.1 mol/L of Tris-HCL with a pH of 8 and a volume 4 times of the lysate is added, the vortexing vibrating is conducted for 10-15 s, and centrifuging is conducted to obtain a supernatant, a DNA solution; and a formula of the lysate is as follows: 0.5 mol/L NaOH and 1% TritonX-100. The method is combined with qPCR to achieve the high-throughput rapid molecular detection of the citrus yellow shoot diseases, and provides a favorable tool for large-scale and accurate diagnosis of the citrus yellow shoot diseases.

The invention discloses a detection kit for a budesonide metabolism marker and a detection method and application thereof. The kit designs a specific amplification primer and a sequencing primer aiming at the polymorphism of GLCCI1A-1106G, and the kit comprises the following components: a sample treating fluid, magnetic beads, an amplification reagent 1, an amplification reagent 2, a GLCCI1A-1106G sequencing primer and a positive control. The detection kit is mainly optimized from two aspects: on one hand, a mode of extracting and amplifying in the same tube is adopted, so that the risk of nucleic acid loss due to repeated tube transfer during extraction is avoided; on the other hand, rapid isothermal amplification is carried out by adding an anti-inhibitor; the invention aims to detect the gene polymorphism of the curative effect of budesonide by combining rapid DNA (deoxyribonucleic acid) preparation, constant-temperature PCR (polymerase chain reaction) amplification and pyrosequencing technologies, and provides suggestions from the gene perspective for clinical personalized medication.

The invention discloses a rapid DNA (deoxyribonucleic acid) extraction kit with magnetic beads for fluorescent quantitative PCR (polymerase chain reaction) detection and an extraction method thereof, and belongs to the technical field of bioengineering. The rapid DNA extraction kit comprises a rapid DNA extraction kit body; the rapid DNA extraction kit comprises a lysis binding solution LB, a washing solution W1, a washing solution W2, a nucleic acid elution reaction solution EB, magnetic beads MB and protease K. The extraction method comprises a manual nucleic acid extraction method or a method for realizing high-flux automatic DNA extraction by matching with an automatic instrument. The rapid DNA extraction reagent with the magnetic bead amplification function can meet DNA extraction of various sample types such as whole blood, plasma and swabs, an extracted product can be directly applied to fluorescent quantitative PCR amplification detection reaction, separation of magnetic beads and nucleic acid is not needed, downstream detection is not affected, and nucleic acid extraction or purification can be achieved more simply, more sensitively and more rapidly.

The invention belongs to the technical field of identification of traditional Chinese medicines and traditional Chinese medicinal materials, and provides a real-time fluorescent PCR method for identifying the authenticity of a garter snake traditional Chinese medicinal material by adopting specific primers, aiming at solving the problems that the existing garter snake detection method is easy to have a false positive result and cannot be applied to market rapid detection and the like. A specific primer sequence used for identifying the authenticity of the garter snake traditional Chinese medicinal material is shown as SEQ ID NO: 1 and SEQ ID NO: 2; a probe sequence is shown as SEQ ID NO: 3. The identification of the authenticity of samples can be finished through rapid DNA extraction and real-time fluorescent PCR specific amplification. The real-time fluorescent PCR method for identifying the authenticity of the garter snake traditional Chinese medicinal material by adopting the specific primers has the advantages that the primer probe double specificity has higher specificity; the fluorescence detection sensitivity is higher; the detection time of the method is short without an electrophoresis process; a pharmacopoeia detection method needs about 4 hours, and the real-time fluorescent PCR method for identifying the authenticity of the garter snake traditional Chinese medicinalmaterial by adopting the specific primers only needs 1.5 hours; the required instrument and equipment are less, and the method can be applied to a rapid detection vehicle service market, and the like.

The invention discloses a universal method for rapidly quantifying RNA residues in a DNA product. According to the method, RNA is quantitatively detected by combining a specific nuclease with a RNA specific fluorescent dye. The method specifically comprises the following steps: digesting DNA by specific nuclease to reserve RNA, eliminating interference, and then quantitatively detecting RNA residues by using the RNA specific fluorescent dye; or, after the RNA is digested by the specific nuclease, quantitatively detecting the RNA by using the RNA specific fluorescent dye, and calculating a difference value before and after digestion to quantitatively detect the RNA residue. Compared with conventional AGE qualitative detection, the method provided by the invention is quantitative detection,and because the RNA specific fluorescent dye has high sensitivity to RNA and is theoretically less influenced by the length difference of RNA fragments, the method provided by the invention has the advantages of good quantitative linearity, high sensitivity, small RNA fragmentation interference and good accuracy; compared with a conventional RT-qPCR mode to indirectly quantifies the RNA residues,the method directly quantifies the RNA residues, is simple and rapid in experimental process and low in cost; theoretically, the method directly quantifies the RNA with more accuracy, and has low requirements on experimenters.

The invention discloses a method for detecting various microorganisms by an integrated multiple nest type PCR one-step method. The method comprises the steps of performing differentiation designing of a universal primer and a specific primer of NM-PCR based on the characteristics of DNA bar code genes of microorganisms to be detected, establishing an NM-PCR one-step detection system, namely performing parameter optimization by using the genomic DNA of the microorganisms to be detected as a PCR reaction formwork, performing quick DNA extraction on microorganism genomes in samples to be detected, performing a NM-PCR amplification reaction by using the genomic DNA in the samples as a formwork, and performing gel electrophoresis detection on PCR reaction products. The method disclosed by the invention concurrently has specificity of the nest type PCR and high flux of multiplex PCR, two-time PCR processes are completed only through one-step PCR reaction, traditional PCR multi-step operations are avoided, the microbial pollution situations of clinical samples such as environment water sources and foods can be quickly and massively detected, the specificity is high, the flus is high, the detection time can be greatly shortened, and the cost can be greatly reduced.

The invention discloses a rapid and low-cost method for separating a PCR template from chicken blood. The rapid and low-cost method for separating the PCR template from the chicken blood comprises thesteps that A, blood corpuscle nucleuses are separated from a blood sample, specifically, a fresh and frozen anticoagulant chicken blood sample is added into a 1.5 mL EP pipe, and 4000rpm centrifugation is carried out for one minute and supernatant is removed; and 500 [mu]l sterile water is added into the pipe to stand for one minute, 9500rpm centrifugation is carried out for three minutes, and the supernatant is abandoned; B, the corpuscle nucleuse dissociation is carried out, specifically, a 3-4% chelated type ion exchange resin solution shaken up by the 200 [mu]l sterile water is added intosediment by using a spearhead with the cut tip part, after mixing, water bath at 55 DEG C-57 DEG C is carried out for 30 minutes, then, a dry heat meter is placed in, and heating is carried out at 100 DEG C for 8 minutes; and C, PCR template separation is carried out, specifically, 9500rpm centrifugation is carried out for two minutes, and the supernatant is taken as a PCR template. Compared witha template obtained by using an existing traditional DNA extraction method, the PCR template obtained by using the rapid and low-cost method for separating the PCR template from the chicken blood hassame effects for PCR technology detecting, the operation is simpler and easier to operate, the time-consuming is short (within one hour), the cost is low (about 0.31 yuan/sample), and the PCR template is suitable for various rapid DNA molecular detection experiments carried out by using PCR technology.

The invention discloses a rapid LAMP detection method for watermelon fusarium oxysporum. According to the method, a set of sensitive and specific LAMP primers are designed by taking a specific DNA fragment of fusarium oxysporum watermelon specialized type as a target, and a rapid LAMP detection system of the watermelon fusarium oxysporum is established by combining a rapid DNA extraction method. The lower detection limit of the LAMP system on FON-1 genome DNA is 10 pg/[mu]L, and the lower detection limit of the LAMP system on the plant bacterium carrying amount is 5 * 10<2> spores/g. According to the system, only about 90 minutes are needed from DNA extraction to detection result acquisition, and pathogenic bacteria hyphae and plant tissues can be used as detection materials. The reaction result can be distinguished by naked eyes. The method has the characteristics of simplicity in operation, high specificity, high sensitivity, high stability and wide applicability, and can provide technical support for rapid diagnosis of field watermelon fusarium oxysporum.

The invention relates to a dual-multienzyme-mediation-based nucleic acid amplification detection method. The method comprises the following steps: pre-amplification, freeze-dried redissolution, amplified detection and result analyzing. According to the method, a normal-temperature nucleic acid amplification technology is combined with a nest type amplification principle, a plurality of enzymatic reactions such as single-stranded DNA binding protein, DNA polymerase and DNA double-strand-opened enzyme are involved, DNA can be amplified by several million folds or more in 10 to 20 minutes under the constant-temperature condition of 30 DEG C to 45 DEG C, and rapid identification on the DNA can be achieved by being matched with a fluorescence detection technique. The method disclosed by the invention is simple in operation, short in time and low in instrument requirements and is applicable to rapid DNA diagnosis.