15 results about "Rapid dna" patented technology

The invention discloses an SSR molecular marker primer related to leek dormancy as well as a detection method and application of the SSR molecular marker primer. Aiming at the problems that the traditional dormancy character identification of Chinese chives depends on field morphological observation as long as 4 months, the efficiency is low and special molecular markers are lacked, seven pairs of specific SSR primers (SSR005 / 170 / 249 / 325 / 423 / 478 / 610) are developed based on cross-species genome re-sequencing (taking green Chinese onion as a reference genome, and the comparison rate is 18.76%-18.89%) for the first time. The method comprises the following steps: establishing a matched detection system: rapidly extracting DNA (deoxyribonucleic acid) by a CTAB (cetyltrimethyl ammonium bromide) method, optimizing a PCR (polymerase chain reaction) system (20 mu L of DNA containing a template of 30 ng, 0.4 U of Taq enzyme, 0.2 mu M of primer, 2.0 mM of Mg < 2 + >, 0.2 mM of dNTPs, and annealing at 57 DEG C), and performing 8% polyacrylamide gel electrophoresis combined with silver staining development, thereby accurately identifying the dormancy character within 7 days. For example, the SSR423 primer has non-differentiated non-dormancy / dormancy germplasm at 177bp, and the coincidence rate of the detection result of 52 germplasm (such as dormancy germplasm 21-1 and non-dormancy germplasm 22-7) and the field phenotype reaches 100%.

The invention discloses a DNA molecular marker rapid detection system and method based on imaging spectrum light splitting, and belongs to the technical field of molecular biological detection. The system comprises an exciting light array 1, a receiving and transmitting integrated detection light path 2, an optical fiber transmission array 3, a light splitting module 4, spectral data processing software 5 and a push-broom structure 6. The excitation light array 1 realizes excitation of different labeled samples; the receiving and transmitting integrated detection light path 2 reflects and focuses the array exciting light to a sample and converges fluorescence generated by the sample to an optical fiber input end; the optical fiber transmission array 3 realizes single-row multi-sample fluorescence transmission; the light splitting module 4 is used for dispersing exciting light and fluorescent light of multiple samples in a single row; the spectral analysis processing software 5 is used for acquiring and preprocessing data; the push-sweeping mechanism 6 realizes row-by-row detection of multiple rows of samples. According to the technology, high-throughput, high-sensitivity and rapid detection of the DNA molecular marker is realized. The method is suitable for the fields of fake material evidence identification, food and biological safety and the like, and has the advantages of simplicity and convenience in operation, strong anti-interference performance and the like.

The invention relates to the technical field of DNA library establishment and sequencing, and particularly discloses a method for performing full-coverage amplification on a DNA sample, a rapid DNA library establishment method, a sequencing method and application thereof. The amplification method comprises the following steps: carrying out low-temperature denaturation PCR (Polymerase Chain Reaction): contacting a DNA (Deoxyribonucleic Acid) sample with a plurality of groups of pre-amplification primer pairs, carrying out PCR amplification, and overlapping an amplification product of any one group of pre-amplification primer pairs in the plurality of groups of pre-amplification primer pairs and an amplification product of any adjacent group of pre-amplification primer pairs with at least 20 basic groups; the library building method comprises the amplification method. The sequencing method comprises the library building method. According to the sequencing method disclosed by the invention, the sequencing result can be quickly and efficiently obtained with high precision and high sequencing depth.

The application discloses a SSR molecular marker primer related to Chinese chives dormancy and a detection method and application thereof. In view of the problems that traditional Chinese chives dormancy trait identification relies on field morphological observation for 4 months, is inefficient and lacks special molecular markers, 7 specific SSR primers (SSR005 / 170 / 249 / 325 / 423 / 478 / 610) are developed based on cross-species genome resequencing (taking Chinese onion as a reference genome, and the alignment rate is 18.76%-18.89%) for the first time. Through the establishment of a matching detection system: CTAB method for rapid DNA extraction, optimization of the PCR system (20 muL contains 30 ng of template DNA, 0.4 U of Taq enzyme, 0.2 muM of primer, 0.2 mM of Mg 2+ 2.0 mM of dNTPs, 57 DEG C annealing), 8% polyacrylamide gel electrophoresis combined with silver staining development, the dormancy trait can be accurately identified within 7 days. For example, the SSR423 primer has no band at 177 bp, and the non-dormant / dormant germplasm cannot be distinguished. The detection results of 52 germplasms (such as dormancy germplasm 21-1 and non-dormancy germplasm 22-7) are consistent with the field phenotypes at a rate of 100%.

The invention relates to the field of agricultural pest drug resistance detection, and provides a detection primer and method for chilo suppressalis ryanodine receptor I4758M mutation and application. According to the detection primer, a mutation recognition site is designed at the 3'tail end of an inner primer BIP, specific detection is achieved through the key base difference (A / G) of a wild type and a mutant type, and the primer group comprises an outer primer pair F3 / B3, an inner primer pair FIP / BIP and a loop primer pair LoopF / LoopB; the detection method is based on loop-mediated isothermal amplification (LAMP), the genotype is visually interpreted through color change of reaction liquid, a thermal cycler is not needed, and the detection method has the advantages of being high in accuracy and sensitivity, easy to operate, low in cost and the like. The matched kit comprises a rapid DNA extraction assembly and is suitable for field on-site detection. The gene mutation frequency of the specific site of the chilo suppressalis can be rapidly detected, and the technical support is provided for resistance monitoring, early warning and scientific treatment of bisamide insecticides.

The utility model relates to a DNA (deoxyribonucleic acid) rapid crude extraction kit based on a paper base, and belongs to the technical field of DNA extraction devices of paper bases. Comprising a cover plate and a bottom groove; the cover plate is detachably connected with the bottom groove; a plurality of groups of circular hollows and a plurality of groups of circular grooves are formed in the cover plate; the circular hollowed-out parts and the circular grooves are opposite in position and consistent in number on the cover plate; a rectangular groove is formed in the bottom groove; a water-absorbing paper base is mounted in the rectangular groove; and a circular DNA recovery paper substrate is arranged in the circular hollow part and is positioned at the upper part of the water-absorbing paper base. According to the kit, the cover plate and the bottom groove are detachably connected, so that the kit is convenient to carry and store; the kit adopts the design of circular hollows and circular grooves, so that the DNA recovery paper substrate is convenient to place and position, complex operation is not needed, and the DNA extraction efficiency is improved; the water-absorbing paper base is adopted to assist liquid transfer, the efficiency is high, DNA does not need to be repeatedly transferred to recover the paper base, and the paper base is effectively prevented from being broken or deformed.

Disclosed are processes and kits for rapid nucleic acid extraction from a nucleic acid-containing material, such as a bone, tooth or semen sample. For bone and tooth process involves providing the nucleic acid-containing material in a form suitable for nucleic acid extraction, adding a lysis buffer to the nucleic acid-containing material to obtain a mixture, mixing the mixture in a manner equivalent for about 30 seconds or longer and separating the mixture by centrifugation to obtain a liquid supernatant. The liquid supernatant contains the extracted nucleic acids which can be used for analysis including STR profiling by conventional or rapid DNA analysis. For semen the processes and kits involve applying an appropriate amount of sperm disruptive agent.

The application discloses a base recognition method based on semiconductor SERS technology and belongs to the technical field of biomedical detection. The specific method comprises the following steps: step one, synthesis of TiO2 nanoparticles; step two, preparation of a TiO2 / 4-MBA system; and step three, base recognition detection. The TiO2 / MBA system can be used for accurate and rapid DNA base recognition, and the developed TiO2 / MBA system is simple to prepare and has excellent base test performance, and has a high market application prospect. The application not only provides a novel base detection method, but also has high sensitivity and selectivity, can recognize and detect four bases and the non-covalent interaction between the four bases and 4-MBA in two pH environments, and has a simple and rapid operation process. The application not only provides a new method for base analysis, but also provides technical support for early diagnosis of DNA related diseases, genomics research and development of molecular sensors, and has a wide application prospect.

The invention provides a plant leaf DNA rapid extraction kit and application thereof, the plant leaf DNA rapid extraction kit is used for rapidly extracting plant leaf DNA, and the kit comprises a DNA rapid extraction solution, a steel ball and a 96 deep hole plate. The rapid DNA extracting solution comprises 15-25 mmol / L of Tris-HCl, 20-30 mmol / L of NaCl, 0.4-0.6 mmol / L of EDTA (Ethylene Diamine Tetraacetic Acid) and 0.10-0.15% of SDS (Sodium Dodecyl Sulfate), and the pH value is 7.5-8.5. When the rapid DNA extracting solution is used, plant leaves, the rapid DNA extracting solution and steel balls are put into a 96 deep-well plate, are subjected to vibration grinding at 50-70 Hz for 30-70 seconds and are centrifuged at 3500-4500 r / min for 5-15 minutes, and supernatant is taken, namely the rapidly extracted plant DNA. The method is simple and rapid, high-temperature or organic solvent treatment is not needed, plant samples can be treated in a high-throughput mode, the concentration of the extracted DNA can reach 29.8-36.9 ng / mu L, and the extracted DNA is suitable for high-throughput genetic typing technologies such as fluorescent quantitative PCR, fluorescence labeled capillary electrophoresis multiple PCR detection and KASP, and has wide application prospects in the fields of crop variety detection, molecular assisted breeding and the like.

The invention relates to the technical field of borosilicate glass, in particular to a sample DNA (deoxyribonucleic acid) rapid extraction method which comprises the following steps: step S1, sample pretreatment: taking 20-100 mu L of liquid sample or 25-50 mg of solid sample, adding 100-200 mu L of pretreatment liquid with the pH value of 7.5-8.0, and uniformly mixing by vortex for 10-30 seconds; s2, synergistic cracking and adsorption: adding 150-200 [mu] L of a composite cracking solution and 10-20 [mu] g of modified diatomite into the pretreatment solution, incubating at 55-60 DEG C for 3-4 min, and reversing and uniformly mixing twice during incubation; s3, impurity removal: adding 150-200 [mu] L of a washing liquid I, standing at room temperature for 1 min, and pouring to remove a supernatant; then adding 200 [mu] L of a washing liquid II, standing for 30 s, and dumping to remove the supernatant. By optimizing the composite lysate, the modified diatomite and reaction conditions, rapid DNA extraction of which the whole process is less than or equal to 8 minutes without protease K digestion and high-speed centrifugation is realized, the method has wide sample adaptability, high automation compatibility and stable DNA yield and purity, the extraction efficiency is greatly improved, and the operation cost and threshold are reduced.

The invention discloses a DNA (deoxyribonucleic acid) rapid extraction method suitable for SSR (simple sequence repeat) and SNP (single nucleotide polymorphism) marker detection of brassicaceous vegetables and application of the DNA rapid extraction method, and belongs to the field of biological breeding. The rapid DNA extraction method provided by the invention comprises the following steps: placing tissues of low-protein parts of brassicaceous vegetables in a porous DNA extraction plate, adding a DNA extraction solution, sealing, carrying out constant high-temperature treatment, centrifuging, and uniformly mixing, so as to obtain the extracted DNA. According to the DNA rapid extraction method suitable for brassicaceous vegetable SSR and SNP marker detection, the to-be-detected sample is directly placed in the porous DNA extraction plate, the DNA extraction process is completed in the same plate, and a tedious tube transferring or liquid transferring step is not needed. According to the whole extraction method, DNA extraction can be completed only through sampling, one-time liquid adding, one-time heating and one-time centrifuging, the operation process only needs 10-15 min, and DNA can completely meet the follow-up PCR amplification requirements of SSR markers and SNP markers.

The invention discloses a DNA fragment rapid assembly method based on programmable nuclease MfAgo, and belongs to the field of molecular biology, the nuclease MfAgo from Methanocaldococcus services is utilized, site-specific cleavage of plasmids can be achieved, and rapid assembly and cloning of cleavage fragments are achieved through T5 exonuclease. Fragment cutting can be completed without adding polymerase and other complex components, the visualization effect is good, and the assembly result can be quickly seen.

The present disclosure relates to improved methods for preparing, sequencing and analyzing short DNA fragments using handheld, nanopore-based sequencing technology as well as improved methods for extracting DNA, in particular genomic DNA, for any downstream application.

The invention relates to a preparation method of an MOFs (Metal-Organic Frameworks) coating microneedle. The preparation method comprises the following steps: preparing a PDMS (Polydimethylsiloxane) female mold with a microneedle model area; preparing a polystyrene polymer solution by using N, N-dimethylformamide, pouring the polystyrene polymer solution to a microneedle model area, and drying and demolding to obtain a polystyrene microneedle; preparing an MOFs precursor solution; a polystyrene microneedle is inversely arranged on the liquid level of the MOFs precursor solution, the solution is kept still for 3-7 days under the room temperature condition, and a single layer of compact and uniform MOFs coating grows on the surface of the polystyrene microneedle in situ; and taking out, washing and airing to obtain the MOFs coating microneedle with the in-situ growth MOFs coating. The method disclosed by the invention has specific enrichment and extraction capability on DNA and can be repeatedly used, and a rapid DNA extraction kit is developed to realize repeatable extraction of DNA in DNA liquid or plant samples.

Compositions, devices and methods for performing an Ultra Rapid Droplet Digital Polymerase Chain Reaction (UR-ddPCR) are provided, for detecting and quantifying target nucleic acid obtained from a sample. The disclosed UR-ddPCR method includes an optional ultra-rapid DNA / RNA extraction method that is compatible with ddPCR and an ultra-rapid thermal cycling (UR-TC) method, the combination of which reduces the tissue-to-result time by about 83%, to ˜20 mins, preferably, <20 mins, when compared to ˜2 hrs required for standard ddPCR.The thermal cycling time of UR-ddPCR is shortened compared to standard ddPCR to reduce the denaturation time from 30 seconds to about 1 to about 3 seconds and the annealing / extension step from 60 seconds to about 1 to about 10 seconds, eliminating final the 10-minute heat inactivation step used in standard commercial (Bio-Rad) ddPCR reactions.