495 results about "Genomic DNA" patented technology

The invention discloses an indel molecular marker for identifying color and luster of citrus peel in a seedling stage and application of the indel molecular marker. The corresponding sites of the indel molecular marker of the citrus are positioned in chr7: 18, 398, 798-18, 399 and 289 intervals of a chromosome 7 of an SWO3.0 reference genome; the genome DNA of the citrus seedling stage plant is used as a template, the Indel molecular marker is amplified through PCR, the citrus seedling stage plant can be divided into a homozygous insertion type, a homozygous deletion type and a heterozygous type according to a PCR amplification result, and the three types are associated with the color of the citrus fruit for identifying the color of the citrus peel, so that the early screening of the seedling stage target traits of the citrus breeding population is realized, and the early screening of the seedling stage target traits of the citrus breeding population is realized. The breeding period is remarkably shortened, the breeding cost is reduced, and important technical support is provided for citrus fruit color improvement breeding.

The invention discloses a method for identifying Yushan black pigs by adopting an SNP (Single Nucleotide Polymorphism) molecular marker technology, and particularly relates to the technical field of molecular detection.The method comprises the following steps: S1, sample preparation: collecting any one of ear tissues, tail tips or blood samples of to-be-detected pig individuals, and extracting to obtain genome DNA (Deoxyribose Nucleic Acid); the genome DNA is used as a template, amplification detection is conducted through a first primer pair, a second primer pair, a third primer pair and a fourth primer pair corresponding to the first SNP site, the second SNP site and the fourth SNP site, and the nucleotide sequences of the first primer pair, the second primer pair and the fourth primer pair are sequentially SEQ ID NO.5-SEQ ID NO.12. By detecting the specific single nucleotide polymorphic sites in the Yushan black pig genome and carrying out genetic typing comparison, the Yushan black pigs and the filial generation thereof can be quickly and accurately identified.

An engineered producer cell comprising an inactivating mutation in one or more endogenous REP binding sites is provided, as are methods for producing the engineered producer cell and using the engineered producer cell to produce a recombinant viral vector and reduce producer cell genomic DNA contamination of a recombinant adeno-associated virus vector preparation.

The invention provides a kit for detecting TP53 gene mutation. The kit for detecting TP53 gene mutation provided by the invention can accurately detect six TP53 gene mutation sites, namely R175H, R249S, R282W, R248Q, R273H and G245S, in genome DNA to be detected at the same time through a multiple fluorescent PCR reaction system and a multi-channel fluorescent signal, can accurately detect the six mutation sites, has ultrahigh amplification efficiency, detection efficiency, sensitivity and specificity, and can be widely applied to detection of TP53 gene mutation. The genotype of a sample with the genome DNA content of 1 ng can be accurately detected, the operation is simple and convenient, and the cost is low.

The present disclosure provides systems and methods to detect somatic or germline variants by providing a predetermined genomic DNA (gDNA) to an assay mixture, and capturing a sample of a subject's genetic information using a DNA sequencer and detecting genetic variants from the genetic information. A mutation may then be classified as being from a germline source if gDNA derived molecules have lengths inconsistent with those expected from cell-free DNA (cfDNA) derived molecules.

The invention relates to a method for extracting genome DNA of plants rich in secondary metabolites and a buffer solution, and belongs to the technical field of molecular biology and botany. The method solves the technical problems of low DNA extraction efficiency, poor purity, easy degradation and the like when a traditional DNA extraction method is used for treating plant tissues rich in secondary metabolites such as alkaloid, polyphenol and the like. Comprising the following steps: adding a complexing agent such as polyethylene glycol or polyvinylpyrrolidone when grinding plant tissues in a liquid nitrogen environment; splitting by using a cell wall splitting buffer solution containing dithiothreitol and a nonionic surfactant; carrying out DNA release and extraction at 60-70 DEG C by adopting a CTAB (Cetyltrimethyl Ammonium Bromide) extraction buffer solution containing polyethylene glycol and papain; and then purifying and precipitating to obtain high-purity genome DNA (Deoxyribose Nucleic Acid). The method can effectively remove alkaloid, protein and other impurities, significantly improves the DNA yield and purity, and is suitable for genome sequencing, genetic resource protection, medicinal plant molecular identification and the like of plants with high secondary metabolites such as Stephania kwangsiensis and the like.

The invention relates to the technical field of biological detection, and discloses a method for double-ring detection of lung cancer L858R mutation based on CRISPR / Cas9 and DNA, and the method comprises the following steps: enriching circulating tumor cells CTC from a blood sample; extracting genomic DNA (deoxyribonucleic acid) of the enriched CTC; carrying out amplification on the DNA fragment containing the L858R mutation by adopting recombinase polymerase amplification RPA (recombinase polymerase amplification); the amplified DNA fragment is hybridized with a recognition ring in a DNA double-ring structure, a DNA double ring is composed of the recognition ring and a report ring, and a part of region of the recognition ring is hybridized with a corresponding fragment of the report ring to form a staggered double-chain region; when L858R mutation exists, the mutated DNA and a recognition ring are hybridized to activate a CRISPR / Cas9 system, and under the guidance of sgRNA, Cas9 cuts hybrid double strands and releases a report ring; carrying out rolling circle amplification RCA under the action of DNA polymerase by taking the released report ring as a template; a fluorescent probe is used for detecting an RCA amplification product, and whether L858R mutation exists or not is judged according to a fluorescence signal. The CRISPR / Cas9 system is combined with a DNA double-ring structure, so that the ultra-sensitive and specific detection on the lung cancer L858R mutation is realized.

The present application relates to a method and system for inferring the age of an individual from semen or a semen stain, and a methylation detection kit, which comprises the following steps: firstly, extracting the genomic DNA of the semen or semen stain of the individual; secondly, performing bisulfite treatment on the genomic DNA; thirdly, selecting a plurality of sites in the CpG sites in the genomic DNA and amplifying the selected CpG sites to obtain the methylation rate of the selected CpG sites; and finally, performing regression analysis on the methylation rate of the selected CpG sites and the age of the individual to construct a regression model for inferring the age of an unknown individual from semen or a semen stain. The average absolute error of the present application reaches 1.68 to 4.44 years.

In order to control plant diseases caused by Xanthomonas bacteria, a novel bacteriophage exhibiting bacteriolytic activity specific to the same bacteria is isolated, and a plant disease control composition containing the same as an active ingredient is developed and provided. Provided are a lysozyme containing a bacteriophage having a novel genomic DNA sequence and exhibiting a lysolytic activity specific to Xanthomonas bacteria, and a plant disease control composition containing the lysozyme as an active ingredient.

Compositions and methods for detecting structural variations (SVs) in target genes or for genetic mapping of movable transposable elements are disclosed, the target genes relating to disease pathologies commonly found in large Mendelian genomics projects, and the movable transposable elements relating to genetic diseases, cancer, and aging. The method comprises: (i) contacting a sample containing genomic DNA with a DNA endonuclease for an effective amount of time to cleave the genomic DNA into fragments, the genomic DNA being uncrosslinked; (ii) subjecting the fragments obtained from step (b) to a DNA ligase to obtain circularized DNA; (iii) subjecting the circularized DNA to reverse PCR amplification containing a reverse primer, wherein the reverse primer is designed to match a expected wild-type sequence near a suspected mutant locus in the gene; and (iv) sequencing the amplified products. Exemplary conditions include Bardet-Biedel syndrome; severe upper and lower limb defects; retinitis pigmentosa; syndromic microcephaly; spastic paraplegia; and atypical hemolytic uremic syndrome.

Described herein are compositions and methods for reducing adaptered-tag sequencing reads during the identification and nomination of on- and off-target CRISPR edited sites. One embodiment is a method for reducing adaptered-tag sequencing reads during the identification and nomination of on- and off-target CRISPR edited sites, the method comprising: contacting in an amplification reaction one or more adaptered-tag blocking oligonucleotides with an isolated genomic DNA having one or more tag sequences and adapter sequences; wherein the adaptered-tag blocking oligonucleotides comprise one or more blocking moieties and hybridize to adaptered-tag sequences at a junction region between the adapter and tag sequences to reduce amplification of the adaptered-tag sequences.

The application relates to the technical field of Chinese soft-shelled turtles, in particular to primers, a kit and a method for identifying the genders of Chinese soft-shelled turtles and application. The genomic DNA of male and female individuals of Chinese soft-shelled turtles is respectively subjected to PCR amplification by using the primer pair, and male-specific fragments and female-specific fragments can be obtained. The primer pair, the kit and the method provided in the embodiments can effectively distinguish female individuals and male individuals of Chinese soft-shelled turtles.

A novel bacteriophage having bacteriolytic activity against bacteria of the genus Salmonella, such as S. Enteritidis, and a bacteriolytic agent consisting of the same are provided. The bacteriophage and the bacteriolytic agent including the bacteriophage have a wide host range for bacteria of the genus Salmonella. A host-specific bacteriophage and an effective Salmonella bacteriolytic agent consisting of the same are provided. A bacteriophage capable of effectively controlling S. Typhimurium, and in particular, multidrug-resistant S. Typhimurium, and a bacteriolytic agent consisting of the same are also provided. A bacteriophage having a specific genomic DNA sequence; a Salmonella bacteriolytic agent consisting of the bacteriophage; and a composition comprising the same are provided.

The application belongs to the field of fish development, and particularly relates to a molecular marker for identifying Nibea albiflora sperm-induced Pseudosciaena heteroclita gynogenesis offspring and hybrid offspring, a primer set thereof, a kit and application. The primer set for identifying the molecular marker of the Nibea albiflora sperm-induced Pseudosciaena heteroclita gynogenesis offspring and hybrid offspring comprises primers F and R, the nucleotide sequence of the primer F is shown as SEQ ID NO. 1, and the nucleotide sequence of the primer R is shown as SEQ ID NO. 2. Based on genomic data analysis, the application develops the molecular marker F / R which can effectively identify the Pseudosciaena heteroclita gynogenesis offspring and hybrid offspring in the juvenile stage. The method comprises extracting genomic DNA of parents and offspring, PCR amplification and agarose gel electrophoresis detection, and finally distinguishing the gynogenesis individuals and hybrid individuals by whether the Nibea albiflora parent marker is contained in the electrophoresis band.

This application relates to a method and primer pairs for identifying allotetraploids from intergeneric hybridization of Chinese cabbage and arugula based on KASP technology. The method includes obtaining a sample to be identified, wherein the sample is obtained by intergeneric hybridization of Chinese cabbage '72049' as the female parent and arugula 'ASTRO' as the male parent, followed by embryo rescue and chromosome doubling; using the genomic DNA of the material as a template, amplification is performed using KASP primer pairs; fluorescence signal detection is performed on the amplified product; the authenticity of the allotetraploid is determined; and distant hybrids can be predicted based on the genotyping results, significantly improving identification efficiency and saving costs.

The invention belongs to the technical field of biology, and particularly relates to a construction method of epigenome edited zebrafish, sgRNA of a targeted zebrafish runx2b gene promoter region and application of the sgRNA. The invention provides a construction method of epigenome edited zebrafish, which comprises the step of carrying out methylation editing by using a CRISPR / dCas9-methyltransferase system targeting runx2b gene promoter region, thereby inhibiting runx2b gene expression and reducing intermuscular bone formation. According to the method, the expression level of the runx2b gene is accurately regulated and controlled through an epigenetic editing technology on the premise that a DNA sequence is not changed, and formation of fish intermuscular bones is effectively reduced. The zebra fish F0 treated by the method represents that the runx2b mRNA expression level is obviously reduced, and the intermuscular bones of the eleventh to sixteenth sarcomedo regions of adult F0 generation fish are obviously shorter than those of a control group, and the shortening range reaches 23.3% to 38.8%. Besides, the method provides theoretical basis and technical support for improving fish economic traits by accurately regulating and controlling the expression mode of the key economic trait gene and simultaneously keeping the integrity of genome DNA (Deoxyribose Nucleic Acid).

The invention discloses an SSR (Simple Sequence Repeat) primer composition for evaluating genetic diversity of channel catfish breeding population and an application method of the SSR primer composition, and belongs to the technical field of fish genetic breeding. The method comprises the following steps: modifying 5'ends of forward primers of the primers by using FAM fluorophores respectively, amplifying genome DNA of each breeding group by using the 10 pairs of microsatellite site primers respectively, analyzing amplification products, reading genotypes, analyzing parameters, and determining the genotype of the breeding group. The analysis result is used for evaluating the population polymorphism, the variation condition of the population genetic structure and the genetic differentiation degree among the populations. By applying the evaluation method disclosed by the invention, the population genetic diversity and genetic structure of continuous breeding generations of channel catfish can be systematically evaluated. The SSR primer and the genetic diversity evaluation method can be used for genetic improvement and new variety breeding of channel catfish.

The application discloses a bacteriocin synthesis regulation gene plnC overexpression and a construction method and application of a recombinant plant lactis bacillus, and has the characteristics that the nucleotide sequence is shown in SEQ ID No:1 in the sequence table, and the construction method steps are as follows: 1) taking the plant lactis bacillus genomic DNA as a template, designing specific primers and performing PCR amplification; 2) purifying and double enzyme cutting the plnC gene fragment, and connecting the linear carrier pMG36e which is also double enzyme cut to obtain a pMG36e-plnC recombination cloning carrier; (3) through an electric transformation method, the pMG36e-plnC is transferred into plant lactis bacillus B1 competent cells to obtain a plnC overexpression strain B1-plnC, and the advantages are that the synthesis of the plant lactis bacillins is promoted, the plnC gene expression amount of the overexpression strain B1-plnC is increased by more than 4.5 times, and the PlnC protein content is increased by more than 1.5 times.

The invention relates to the technical field of nucleic acid detection, and discloses a method and device for detecting and distinguishing radix trichosanthis, poria cocos and radix puerariae and terminal device.The method comprises the steps that genome DNA of a to-be-detected sample is extracted, a specific DNA fragment is amplified through a universal amplification primer and a specific amplification primer, and an amplification product is obtained; the method comprises the following steps: respectively constructing independent detection modules aiming at radix trichosanthis, poria cocos and radix puerariae, wherein each independent detection module comprises Cas12a protein, a test probe and specific crRNA; the specific crRNA is obtained by screening according to a PAM sequence and a detection contrast ratio. According to the invention, genome DNA of a to-be-detected sample is extracted, a universal amplification primer and a specific amplification primer are utilized, then an amplification product is added into an independent detection module, and specific crRNA is utilized to carry out specific recognition on the amplification product, so that a molecular detection system aiming at radix trichosanthis, poria cocos and radix puerariae is established, and effective distinguishing of different traditional Chinese medicinal material components is realized.

The present invention is directed to improved materials and methods related to cytosine and adenosine base editing of genomic DNA.

The invention relates to a method of determining somatic allele-specific copy number alterations (CNAs) in the genomes of cells in a test-sample from a subject, the method comprising: i) providing an indexed-DNA library of DNA fragments resulting from whole-genome amplification of genomic DNA from cells of the test-sample, ii) providing whole genome sequencing data of reference non-cancer cells from a reference-sample from the subject; and iii) determining somatic allele-specific copy number alterations in the genome(s) of the cells of the test-sample; and associated methods and uses in cancer therapy.

The application discloses a molecular marker, primer, method and application for identifying tobacco root rot Fusarium proliferatum, and belongs to the technical field of plant disease molecular detection. The molecular marker is a nucleotide sequence shown in SEQ ID NO. 1, and can accurately distinguish tobacco root rot Fusarium proliferatum from its close species, i.e., Fusarium solani, Fusarium equiseti and Fusarium oxysporum. The specific primer pair based on the marker is composed of FPITS-3F (5'-GCTTGCCGCAAGGGCT-3') and FPITS-3R (5'-AAGGTCTCCGTTGGTGAACC-3'). The target bacteria can be rapidly identified by using the primer pair to perform PCR amplification and detecting whether a 396 bp characteristic band exists in the amplification product. The primer has high specificity and has no cross reaction to the above close species. The sensitivity is high, and the lower limit of detection of genomic DNA is 1*10-3 ng*muL-1, and the spore detection sensitivity in soil is 1 CFU / g. The method is simple in operation, and the detection can be completed in 2 hours only by using a common PCR instrument, and is suitable for early rapid diagnosis, monitoring and early warning in laboratories and production lines.

The present disclosure generally relates to methods and compositions for interrogating and / or analyzing DNA methylation in a biological sample. In some aspects, the present disclosure relates to methods for determining the methylation status of a region of interest of genomic DNA. In some aspects, the methylation status is analyzed by interrogating converted DNA in which the sequence of the converted DNA is indicative of the methylation state of the DNA. In some aspects, the methods comprise generating a collective signal that is based on the methylation states of a plurality of sequences or residues in the DNA, and that is representative of the methylation status of the region of interest as a whole.

The application discloses a method for improving chromatin DNA accessibility in cells and application thereof, relates to the technical field of gene sequencing and tissue cell sample analysis, and comprises the following steps: providing a sample to be detected; mixing the sample to be detected with an alcohol reagent to permeabilize cells and expose heterochromatin regions of the sample to be detected, so as to obtain a pretreated sample; performing fragmentation treatment on DNA of the pretreated sample, performing reverse transcription reaction on RNA in the pretreated sample, performing labeling on the fragmented DNA and the cDNA after the reverse transcription reaction, and separating labeled genomic DNA and cDNA; and performing library construction and sequencing on the fragments of the labeled genomic DNA and the cDNA product respectively, so as to obtain single-cell genomic information and transcriptome information of the sample to be detected. The alcohol reagent can open the heterochromatin without bias to obtain more nucleic acid fragments, improve the sensitivity of single-cell genome detection, does not affect RNA detection, and realizes high-throughput double-omics analysis of single-cell whole-genome DNA and RNA.

The present application provides a primer pair for detecting bovine genomic DNA, a detection reagent or kit comprising the primer pair of the present application, and a method for detecting bovine genomic DNA using the primer pair, wherein the primer pair specifically binds to the sequence shown in SEQ ID NO: 1. The PCR detection method using the primer pair is simple and fast to operate, has high sensitivity, and can distinguish interfering DNAs such as pig, CHO, Vero, human, NS0, MDCK, E. coli, Pichia and Sf9.

The application relates to the technical field of Chinese soft-shelled turtles, in particular to primers, a kit and a method for identifying the genders of Chinese soft-shelled turtles and application. The genomic DNA of male and female individuals of Chinese soft-shelled turtles is respectively subjected to PCR amplification by using the primer pair, and male-specific fragments and female-specific fragments can be obtained. The primer pair, the kit and the method provided in the embodiments can effectively distinguish female individuals and male individuals of Chinese soft-shelled turtles.

This invention relates to molecular marker 1769 and its application in the identification of sex-reversed pseudomale individuals in the Northeast Forest Frog (Rana davidii), and pertains to the field of molecular markers. To address the current difficulty in identifying sex-reversed pseudomale individuals in the Northeast Forest Frog, this invention extracts the genome from the muscle tissue of the left hind limb of the Northeast Forest Frog. Using the extracted genomic DNA as a template, PCR amplification is performed using primer pairs containing the sequences shown in SEQ ID NO:1 and SEQ ID NO:2. The PCR amplification products are detected by electrophoresis on a 3% agarose gel. Phenotypic male individuals exhibiting the 1769 bp band corresponding to the nucleotide sequence shown in SEQ ID NO:3 are genetically male Northeast Forest Frogs, i.e., true males. Phenotypic male individuals lacking the band corresponding to the nucleotide sequence shown in SEQ ID NO:3 are pseudomale individuals who have undergone sex reversal from genetically female to phenotypically male. Phenotypic female individuals lacking the band corresponding to the nucleotide sequence shown in SEQ ID NO:3 are female individuals.

The invention belongs to the technical field of microbiological detection, and particularly relates to a rapid detection method for listeria monocytogenes RAA-CRISPR / Cas12a and application of the rapid detection method for the listeria monocytogenes RAA-CRISPR / Cas12a. According to the present invention, based on the combination of the recombinase polymerase amplification (RAA) and the CRISPR-Cas12a system, the ultra-sensitive and rapid detection of the Listeria monocytogenes is achieved through the constant temperature nucleic acid amplification and the specific gene recognition; the sensitivity reaches 3.6 CFU / mL (bacterial liquid) and 3.78 copies / L (genome DNA); the detection time is shortened to 26 minutes, and the method can be applied to rapid detection of various environmental samples.

Provided according to one aspect of the present disclosure is a composition for wound healing or tissue regeneration that comprises a genomic DNA fragment extracted from human placenta-derived mesenchymal stem cells and a placental growth factor and which exhibits excellent tissue regeneration ability. Provided according to another aspect of the present disclosure is a preparation method which enables the efficient, biocompatible, economical, and environmentally friendly production of a composition for wound healing or tissue regeneration having excellent tissue regeneration ability.

The invention provides a method for screening candidate genes and SNP (Single Nucleotide Polymorphism) loci related to residual feed intake of Sahu hybrid sheep, which is characterized by comprising the following steps: S1, collecting jugular vein blood samples of the Sahu hybrid sheep, and extracting genomic DNA (Deoxyribose Nucleic Acid) of the blood samples for quality detection; the method comprises the following steps: S1, extracting DNA, S2, carrying out whole genome re-sequencing on the extracted DNA and carrying out genotyping to obtain SNP genotype data, and S3, carrying out reference genome comparison, SNP detection and genotype quality control. And S4, carrying out whole genome association analysis on the residual feed intake character of the Sahu hybrid sheep to obtain a significant SNP site. The nucleotide sequence of the SNP site obviously related to the residual feed intake of the Sahu hybrid sheep, which is obtained by the method provided by the invention, is as shown in SEQ ID NO.1, the basic group R at the 51st site of the sequence is A or G, the gene mutation causes the nucleotide of the sequence to generate polymorphism, and when the marker is mutated into G, the Sahu hybrid sheep shows lower residual feed intake.