25 results about "Dna concentration" patented technology

The invention discloses a specific detection target PsRrp8 of phytophthora sojae, a DNA (deoxyribonucleic acid) sequence of the specific detection target PsRrp8 is shown as SEQ ID NO.1. The invention further provides a primer combination for detecting the phytophthora sojae, the primer combination comprises a forward primer and a reverse primer, the sequence of the forward primer PsRrp8-qPCR-F1 is shown as SEQ ID NO.2, and the sequence of the reverse primer PsRrp8-qPCR-R1 is shown as SEQ ID NO.3. The invention further provides a kit for detecting the phytophthora sojae. The lowest detectable DNA concentration of the detection primer combination provided by the invention on the phytophthora sojae (P.sojae) is 0.332 pg.L <-1 >, which indicates that the quantitative detection technology has high sensitivity. The method can be successfully applied to phytophthora detection of phytophthora root rot disease field plants, phytophthora sojae can be rapidly detected, and a new technical means is provided for early warning of diseases.

The present invention is directed to a method of determining DNA concentration in a DNA virus. The invention features three basic steps. The initial step is the capture of a defined amount of virus capsid particles on a first solid phase. The second step is lysis of the capsid to release the virus DNA from the first solid phase into a lysis solution. After separating the lysis solution from the first solid phase, the third step is contacting the lysis solution with a second solid phase. The second solid phase captures total DNA derived from the captured capsid. The present invention is also directed to a method for measuring the percentage of full virus capsid, comprising first determining the ssDNA concentration in viruses, and then converting the ssDNA concentration to percentage of full virus capsid using a calibration curve having DNA concentration plotted against standards of % of full capsids.

A method for extracting DNA from Kappa longicornis, relating to the field of molecular biology, includes the following steps: grinding a Kappa longicornis sample into powder; adding lysis buffer preheated to 60–65°C to the powder and incubating at 60–65°C; adding Proteinase K for protein digestion; adding RNase A for RNA digestion; extracting with a mixture of chloroform and isoamyl alcohol, centrifuging, and collecting the supernatant; adding pre-cooled isopropanol to the supernatant for precipitation, centrifuging, and obtaining the DNA precipitate; washing and drying the DNA precipitate; and dissolving the DNA in TE buffer or nuclease-free water. The beneficial effects of this invention are as follows: by optimizing the lysis buffer and combining it with stepwise enzymatic hydrolysis and organic extraction processes, polysaccharides and pigments of *Kappa longicornis* can be efficiently removed, and the extracted DNA concentration is >300 ng / μL, with a purity of A260 / A280≈1.76 and A260 / A230≈2.21, meeting the requirements of downstream molecular experiments; at the same time, reduced glutathione is used instead of 2-mercaptoethanol, making the operation safe and odorless, with low reagent costs, and suitable for batch sample processing.

The invention provides a primer, a probe and a kit for identifying coreius guichenoti based on environmental DNA, and a specific primer and a probe designed according to a COI gene sequence in coreius guichenoti mtDNA, namely the primer and the probe can specifically identify coreius guichenoti in numerous fishes, so that the detection time is greatly shortened, the detection can be completed within 6 hours, and the detection efficiency is greatly improved. The method is simple in operation, accurate in monitoring result and high in sensitivity, the lowest DNA concentration of real-time fluorescence PCR detection can reach the pg level, and the method better meets the actual requirements of eDNA field detection in the upstream of the Yangtze River.

The invention belongs to the technical field of detection devices, and discloses a detection device and a rapid detection method for enteromorpha microscopic propagules. According to the detection device, a connector and a sample feeding pipe and a connecting pipe at the bottom and the top of a filter are limited through a first protection assembly, a second protection assembly and a limiting assembly respectively, and meanwhile, protection blocks are used for protecting the outer sides of the first protection assembly, the second protection assembly and the filter, so that the filter is prevented from being easily loosened due to vibration during operation of the filter; the rapid detection method comprises the following steps: 1, filtering a collected water sample through a filter membrane in a filter instrument, and extracting high-purity DNA (deoxyribonucleic acid); 2, cloning and purifying the enteromorpha specific gene, determining the DNA concentration of the target gene, and establishing a standard curve; and 3, detecting the copy number of the enteromorpha microcosmic propagule in the water body through qPCR, and calculating the cell abundance according to the copy number of the target gene in a single cell to realize rapid identification and quantitative detection.

The invention discloses a dual digital PCR (polymerase chain reaction) detection method for monkey campylobacter jejuni and monkey enterocolitis yersinia, and relates to the technical field of microbiological detection, the detection method comprises the following specific steps: multi-source heterogeneous data collection: collecting monkey feces and intestinal tissue samples, extracting genome DNA (deoxyribonucleic acid) by using a phenol-chloroform method, detecting the concentration and purity of DNA, and detecting the concentration and purity of the DNA; after reaching the standard, storing at-20 DEG C for later use; according to the method, high-sensitivity and specific detection on the two pathogenic bacteria is realized through a combination of well-designed primers and probes, according to the method, a gyrA gene conserved region of monkey campylobacter jejuni CJ and a POL gene conserved region of monkey yersinia enterocolitica YE are used as detection targets, and specific fragments of 249bp and 182bp are amplified respectively; the method effectively avoids interference of non-specific amplification, can simultaneously detect two pathogenic bacteria in a single reaction system through a dual digital PCR technology, accurately calculates target gene copy number concentration through a Poisson distribution formula, and improves detection accuracy and reliability.

The application discloses a kind of construction methods of matrix solver based on DNA computing, it is related to molecular computing technology field, comprising the following steps: obtaining matrix to be calculated, the element of the matrix to be calculated is converted into DNA concentration difference signal and DNA circuit signal, generates double-track encoding matrix signal;Based on the double-track encoding matrix signal, construct simple operation module and complex operation module;With chemical reaction network, the output concentration difference of the simple operation module and the output concentration difference of the complex operation module are uniformly operated, and a steady-state output signal is generated;The steady-state output signal is subjected to weight calibration and calculation result convergence, and a matrix solver based on DNA computing is generated.Benefiting from this, the low computational efficiency of the existing optimization and control algorithm is solved, the optimal decision cannot be realized quickly, stably and with high precision, resulting in the path smoothness of robot in actual operation being insufficient, the energy consumption being relatively high, and the environmental adaptability being weak.

The present application belongs to the field of biological detection, and particularly relates to a field visual rapid detection method for citrus Huanglongbing. The present application is based on MIRA-lateral flow test paper, takes the outer membrane protein (OMP) of Asian species of Huanglongbing bacteria of citrus as a target, designs and selects specific primers and probes, and prepares dry powder microspheres together with MIRA reaction enzymes, mixes the field rapid extraction plant DNA lysate, dry powder microspheres and Buffer microspheres in a closed chamber to react, adopts a miniature temperature control device to complete amplification under the condition of 42 DEG C for 20 minutes, and directly realizes visual interpretation of the associated test paper strip. The detection method has no cross reaction with other citrus pathogenic bacteria, the sensitivity test can detect the DNA concentration lower limit of 6.9x10 ‑3 ng / μL, the detection device is small in size and convenient to carry, the whole process does not need professional operation or precise instruments, the detection can be completed within 30 minutes, and the detection method is suitable for rapid detection of citrus Huanglongbing by grass-roots agricultural technicians, planting enterprises and fruit farmers.

This invention discloses a method, system, and device for diagnosing malignant tumors or predicting the treatment efficacy of malignant tumor patients based on the characteristics of mature erythrocyte DNA. This invention investigated the clinical significance of erythrocyte DNA in patients with malignant tumors, finding differences in RBC DNA characteristics between healthy individuals and patients with malignant tumors. Further research revealed a correlation between RBC DNA concentration and treatment efficacy in patients with malignant tumors. Based on this, this invention provides a method for diagnosing malignant tumors based on the characteristics of mature erythrocyte DNA, and also provides a method, system, and device for predicting the treatment efficacy of malignant tumor patients based on erythrocyte DNA concentration. This invention provides a reference for understanding RBC DNA and exploring its potential as a clinical biomarker.

The invention relates to a planet identification method based on a KASP marker. The planet identification method comprises the following steps: (1) carrying out surface disinfection on a plant sample to be tested; (2) grinding the disinfected plant leaves into powder by using liquid nitrogen; (3) extracting the genome DNA of the sample, adjusting the DNA concentration to 5-50ng / mu L, and storing at 4 DEG C for later use; (4) selecting a specific SNP (Single Nucleotide Polymorphism) site on the basis of comparing the ppc-1E1c sequences of the astrophe plants; (5) designing two forward primers and a reverse primer according to the SNP site; (6) respectively adjusting the concentrations of the three primers to 36 [mu] M, 36 [mu] M and 90 [mu] M, and mixing the three primers according to a volume ratio of 1: 1: 1 to form a primer mixed solution; (7) preparing a reaction solution according to a reaction system: 5 [mu] L of template DNA, 5 [mu] L of KASP mastermix and 0.14 [mu] L of the primer mixed solution, and setting a negative control; (8) performing amplification according to a PCR program; and (9) judging whether the sample is a planet plant or not according to a fluorescence typing result. The method is rapid, accurate, low in cost and suitable for species identification, germplasm resource identification and molecular marker-assisted breeding of planets.

The invention discloses a pomacea canaliculata environment DNA specificity detection line and a biomass evaluation method, the technical scheme is as follows: the pomacea canaliculata environment DNA specificity detection line comprises an upstream primer, a downstream primer and a probe, and the method uses the pomacea canaliculata environment DNA specificity detection line to detect the pomacea canaliculata environment DNA concentration and obtain the environment DNA detection quantity; and calculating the biomass of the pomacea canaliculata according to the obtained environmental DNA detection quantity, namely, detecting the environmental DNA concentration of the pomacea canaliculata, and calculating the biomass of the pomacea canaliculata according to the environmental DNA detection quantity by using a pomacea canaliculata biomass calculation formula. The method has the advantages that only the pomacea canaliculata is specifically detected, other freshwater snails cannot be detected wrongly, the problem of wrong detection is effectively solved, particularly, different haplotypes of various pomacea canaliculata are covered, the environmental DNA monitoring work efficiency of the pomacea canaliculata is improved, and the established biomass evaluation model based on environmental DNA degradation and shedding rate has the advantages that the environmental DNA degradation and shedding rate of the pomacea canaliculata can be effectively evaluated. The method can be successfully applied to field biomass evaluation of ampullaria gigas.

Methods of determining the concentration of a nucleic acid of interest in a liquid sample are provided. The method uses a capture oligonucleotide and one or more signal oligonucleotides that hybridize to different regions of the nucleic acid of interest in the liquid phase. The hybridized complexes are then captured by a solid phase by a capture oligonucleotide and detected by one or more signal oligonucleotides. In one embodiment, a Bio-Layer Interferometry (BLI) technique is used to detect DNA concentrations. BLI is an optical technique for measuring macromolecular interactions by analyzing the interference pattern of white light reflected from the surface of the biosensor tip.

The method for rapidly detecting the authenticity of the yak meat comprises the following steps: extracting DNA of a meat product to be detected by using the meat DNA sampler to obtain a DNA template; performing qPCR (quantitative polymerase chain reaction) detection on the obtained DNA template to obtain a Ct value; and comparing the obtained Ct value with a preset qPCR standard curve to obtain the DNA concentration of the meat product to be detected. And judging the authenticity of the meat product to be detected according to the obtained DNA concentration. The method effectively solves the problems of difficulty in field detection of meat adulteration and slow evidence collection for a long time, has the outstanding advantages of simplicity and convenience in operation, reliable result and excellent efficiency, and has remarkable economic and social values for protecting rights and interests of consumers and maintaining fair and order of the market.

The present invention relates in particular to an in vitro method for detecting parasite DNA of the genus Schistosoma from a urine sample of a subject likely to have been infected by said parasite, comprising an isothermal amplification step with a set of pan-specific primers capable of simultaneously amplifying and detecting the species S. haematobium, S. mansoni, S. bovis, S. curassoni, and S. japonicum. It also relates to a simplified in vitro method, adapted to endemic areas, for extracting parasite DNA, particularly of the genus Schistosoma, from a biological sample (e.g., urine) of a subject likely to have been infected by said parasite, comprising steps of DNA concentration by decantation, lysis with heating but without agitation or centrifugation, and filtration and elution of the parasite DNA.

The invention belongs to the technical field of nucleic acid detection, and relates to a nucleic acid detection method and kit by combining digital PCR with surface enhanced Raman scattering (SERS-dPCR). A Raman enhanced substrate with positive charges and a Raman probe coupled with Raman reporter molecules are utilized, positive / negative micro-reaction units are distinguished by detecting significant reduction of Raman signals caused by probe degradation caused by PCR amplification, and the target DNA concentration is calculated according to Poisson distribution. Compared with conventional digital PCR, the SERS-dPCR method provided by the invention has the remarkable advantages of remarkably shortened detection time (for example, only 25 cycles are needed), high sensitivity, accurate quantification, strong multiple detection performance and the like, and is suitable for carrying out rapid, specific and stable absolute quantitative detection on nucleic acid.

The application relates to the technical field of biological detection, and discloses an aflatoxin-producing gene detection method based on magnetic nanoparticles and carbon quantum dots, which comprises the following steps: mixing a to-be-detected sample containing target DNA and MNPs-capture probes, carrying out first incubation to generate base complementary pairing and combination; mixing the formed product and CQDs-signal probes, carrying out second incubation to generate hybridization reaction, so as to form a sandwich structure; separating the reaction system by using an external magnetic field, collecting supernatant after separation; testing the fluorescence intensity of the supernatant, and determining the concentration of the target nucleic acid according to the positive correlation between the fluorescence intensity reduction degree and the target DNA concentration. The application directly sets the fluorescence test object as the supernatant after separation by combining the magnetic separation technology with the double-probe sandwich method, eliminates the dissociation and elution steps of the probes in the conventional method, eliminates the optical interference of food matrix by using magnetic separation, simplifies the operation process, and improves the quantitative detection accuracy.

The invention relates to the technical field of molecular detection of malus micromalus makino, in particular to a malus micromalus makino SS-COI PCR (polymerase chain reaction) detection primer and a detection method. The nucleotide sequence of an upstream primer of the Smith microgramma SS-COI PCR detection primer is as shown in SEQ ID No. 1, and the nucleotide sequence of a downstream primer of the Smith microgramma SS-COI PCR detection primer is as shown in SEQ ID No. 2. Sensitivity detection results show that the detection limit DNA (deoxyribonucleic acid) concentration of the margarine SS-COI PCR detection method provided by the invention is 3.427 * 10 <-2 > ng / mu L, which shows that the detection sensitivity is high.

The invention discloses a nucleic acid aptamer capable of specifically recognizing fusarium oxysporum as well as a screening method and application of the nucleic acid aptamer. The nucleic acid aptamer is at least one of Fu01, Fu02 and Fu03, and the sequences of the nucleic acid aptamer, namely, Fu01, Fu02 and Fu03, are respectively as follows: Fu01: 5 '-TGACCCGTCTAAGCACGGCGTGCTCCTGGAGCACGGTACGGTCA-3' (SEQ ID NO. 1), Fu02: 5 '-ATAGTCCCTGGAGCAGGTACGGTCA-3' (SEQ ID NO. 2), Fu03: 5 '-TGACCAGCAGCAGGGACGATG-3' (SEQ ID NO. 3), and the sequences of the nucleic acid aptamer, namely, Fu01, Fu02 and Fu03 are respectively as follows: 5 '- Fusarium is incubated with the ssDNA library for reverse screening, so that non-specifically bound ssDNA is removed, and the specificity of the aptamer is enhanced. And carrying out high-speed centrifugation to obtain an eluent, precipitating DNA and measuring the DNA concentration. And carrying out ePCR and purification on the DNA to obtain dsDNA required by the next round of screening. And putting the obtained dsDNA into the next round and all subsequent rounds of screening. After ten rounds of screening are completed, a DNA sample is detected, after the DNA sample is qualified, the DNA sample is subjected to terminal repair and A tail addition, sequencing linkers are connected to the two ends of a fragment respectively and purified, PCR amplification is not carried out in the middle, and the PCR-free library is prepared. And carrying out high-throughput double-end sequencing by using an Illumina platform, wherein 150bp is detected at each end.

The invention belongs to the technical field of gene extraction, and particularly relates to a method for increasing the yield of cell genome DNA. Aiming at the technical problem of low yield in trace cell samples in the traditional DNA extraction method, the invention provides a method for washing cells by using a solution containing 1, 6-hexanediol and utilizing the characteristic of depolymerizing a protein aggregate, so that the chromatin structure is loose, and the release efficiency of protease K on genome DNA is remarkably improved. The method is simple and convenient to operate and suitable for adherent or suspension cells. Experiments prove that the DNA concentration is increased by about 67% compared with that of a traditional method, and the purity (A260 / A280) has no significant difference.

This invention discloses specific primers, detection methods, and applications for the detection of tet(X)-positive Acinetobacter amysii. The specific primers include amysii-6712F / amyesii-6712R and ase-F / ase-R. Using these primers, PCR amplification of genomic DNA or single colonies of tet(X)-positive Acinetobacter amysii was performed, with band sizes of 690 bp and 318 bp, respectively. The detection accuracy of tet(X)-positive Acinetobacter amysii was 100%, demonstrating high accuracy. The limit of detection (LOD) DNA concentration was only 0.003 ng / μL, indicating high sensitivity. The total time for colony picking and detection was only about 2.5 hours. For tet(X)-positive Acinetobacter amysii, the method based on specific PCR primers for detection is simple, accurate, and rapid, and has broad application prospects and economic value.

This invention discloses a specific detection target Pc_s376271 for Pythium cedri and its application. The specific detection target includes the nucleotide sequence Pc_s376271 as shown in SEQ ID NO.27. The LAMP primer composition designed based on the above target gene disclosed in this invention can be used for rapid and efficient detection of Pythium cedri. The detection method has the advantages of high specificity, high accuracy, good sensitivity, simple operation, and visually observable results. When the DNA concentration reaches 100 pg·μL... ‑1 It can identify Pythium cedarum in a short time, making it highly practical.

The invention discloses a single plasmid integrated CRISPR (clustered regularly interspaced short palindromic repeats) / Cas9 gene editing system as well as a preparation method and application thereof. According to the invention, the Cas9 nuclease and the gRNA expression cassette are integrated in a single vector, so that saccharomyces cerevisiae cells can be introduced at one time, accurate editing of target genes is realized, the problems of unstable plasmids, difficulty in screening and multiple times of transformation in a traditional double-plasmid system are avoided, and the experimental repeatability and the operation convenience are greatly improved. The sgRNA target sequence, the length of the homologous arm, the donor DNA concentration and the competent preparation condition are systematically optimized, and the editing efficiency can stably reach 90% or above. The yield of rare ginsenoside Rh1 in a modified strain obtained by using the system is improved by 30.4%, the accumulation of precursor PPD is reduced by 51.33%, and the Rh1 synthesis efficiency is remarkably improved.

The application discloses a specific detection target PsRrp8 of soybean phytophthora, a DNA sequence of the specific detection target PsRrp8 is shown as SEQ ID NO. 1, and further provides a primer combination for detecting the soybean phytophthora, the primer combination comprises a forward primer and a reverse primer, a sequence of the forward primer PsRrp8-qPCR-F1 is shown as SEQ ID NO. 2, and a sequence of the reverse primer PsRrp8-qPCR-R1 is shown as SEQ ID NO. 3. P. sojae The minimum DNA concentration of the soybean phytophthora (Phytophthora sojae) detectable by using the detection primer combination provided by the application is 0.332 pg*muL ‑1 , and the quantitative detection technology has high sensitivity. The application can be successfully applied to the detection of the soybean phytophthora in a plant in a phytophthora root rot disease field, can rapidly detect the soybean phytophthora, and provides a new technical means for early warning of diseases.

Fragmentomic signatures provide various characteristics of a sample (e.g., urine or plasma) and / or a subject. Fragmentomic signatures of urinary cell-free DNA can be used to determine the relative contribution or enrichment of clinically relevant DNA (e.g., transrenal and non-transrenal urinary cfDNA types). Such measurements reflect glomerular permeability and can be used to monitor various diseases, such as kidney abnormalities. Fragmentomic signatures can include corrected urinary DNA concentration, size, and terminal motifs of urinary DNA molecules, and cfDNA molecules derived from open chromatin regions (OCRs) of one or more tissues. In addition, nuclease activity or other cfDNA fragmentation processes can be determined based on the relative contribution of different cfDNA cleavage profiles, which can also be used to determine the contribution of tissue-derived cfDNA, the level of pathology, and gestational age.

The application discloses a method for parentage identification, and belongs to the technical field of parentage identification. S101: sequencing of the biallelic loci of the two free DNA samples of the pregnant woman during pregnancy respectively obtains DNA data S1 and S2; S102: respectively obtaining the typing locus set X(S1) and X(S2) containing the fetal signal in S1 and S2; S103: respectively calculating the DNA concentration p1 and p2 of the fetus according to S1 and S2; S104: calculating the common rate K according to formula II, K=[X(S1)∩X(S2)] / [X(S1)∪X(S2)](II); S105: judging the genetic relationship of the fetus during the two pregnancies according to the clustering model of K, p1 and p2. Without a doubtful father sample, whether the fetus is the same father can be judged by comparing the DNA data between the two independent pregnancies.