15 results about "Dna concentration" patented technology

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.

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 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 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 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 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.

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 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.