163 results about "Agarose electrophoresis" patented technology

The invention discloses a detection method for Y chromosome micro-deleted gene. In the method, after human peripheral blood genome DNA is extracted, multiple PCR primer design is used, wherein a multiple PCR reaction system comprises N pairs of chimeric primers for specifically amplifying the STSs site in an AZF segment of a Y chromosome and one pair of universal primers. N+1 pairs of primers are participated in the multiple PCR reaction, and the reactions of DNA template such as denaturalization, anneal and extending in circulating amplification target are carried out in a same reaction system; 10ulPCR products are taken and dye by EB, and electrophoresis is carried out through 3% agarose. The electrophoresis voltage is 4 V/cm, and the electrophoresis time is 20 minutes; and then the electrophoretogram is observed under an ultraviolet transilluminator so that the result is judged. Therefore, a detection method with simplified PCR reaction conditions, high amplification efficiency and easy operation for clinic experiments is established.

The invention belongs to the technical field of biotype identification, in particular to a method for identifying indica type rice and japonica rice using rice grain by using a rice grain (rice) and inserting or deleting (InDel) a molecular marker. The method comprises the following steps of: extracting DNA from the rice grain, and comparing full-genome DNA sequences of indica type rice 93-11 andjaponica rice Nipponbare to obtain 40 pairs of specific InDel primers; and performing fragment amplification, electrophoretic separation and electrophoresis pattern analysis on the extracted DNA the in rice seed to identify the characteristics of the indica type rice and japonica rice of a rice sample. The method concretely comprises the following steps of: taking the DNA extracted from the rice grain as a template, and analyzing and counting molecular fingerprint patterns obtained on the basis of a polymerase chain reaction and agarose electrophoresis by using combination of the 40 pairs of specific InDel primers; and determining the characteristics of the indica type rice and japonica rice of the tested rice seed (sample) according to gene frequency (Fi or Fj) calculated by 93-11 genotype and japonica rice Nipponbare genotype molecular fingerprint on 40 InDel loci of the sample. The result can be obtained by only detecting the sample of one grain of seed, and the method is convenient and rapid, accurate in identification, and has good popularization and application prospect.

The invention discloses a method for inspecting purity of two-line hybrid rice seeds by utilizing an SSR (Simple Sequence Repeat) method, in particular a method for inspecting the purity of Fengliangyou No. 1 rice or Fengliangyouxiang No. 1 rice seeds by utilizing SSR molecular markers. The method comprises the following steps of: performing amplification and agarose electrophoretic separation on genome DNA (Deoxyribonucleic Acid) of the two-line hybrid rice varieties Fengliangyou No. 1 and parents thereof or Fengliangyouxiang No. 1 and parents thereof respectively by utilizing primers P1 and P2 to find out a characteristic spectrum band of difference between a first-filial generation and the parents; inspecting the samples of Fengliangyou No. 1 or the Fengliangyouxiang No. 1 by calculating the purity of the inspected seeds; and comparing and contrasting the inspected purity with purity tested by field planting to confirm that the results of the two are highly consistent. The method is suitable for inspecting the authenticity and variety purity of the Fengliangyou No. 1 or Fengliangyouxiang No. 1 two-line hybrid rice and parents thereof.

The invention relates to a method for detecting salmonella through PCR-DHPLC, belonging to the technical field of bioanalytical chemistry. The method adopts PCR-DHPLC to detect salmonella and comprises the following steps: cultivating salmonella, preparing magnetic nanoparticles, extracting the DNAs of the salmonella genome, selecting the specific primer of salmonella, performing PCR amplification and performing DHPLC detection to the PCR product. The method of the invention adopts DHPLC instead of the traditional agarose electrophoresis to detect the PCR product, thus simplifying the reaction conditions, increasing the detection sensitivity, simplifying the operation and achieving the aim of large flux; and compared with the traditional microbial detection method and the PCR-gel electrophoresis method, the PCR-DHPLC detection method has the advantages that the sensitivity is high, the specificity is high, the detection is fast and accurate, the analysis of result is simple, the demand on samples is not high, the large flux detection can be performed, etc.

The invention belongs to the technical field of molecular biology, which relates to a PCR-based practical flux detection method for molecular markers of the quality characteristics of wheat. In the method, specific molecular markers of NullAx (Glu-A1 locus), Bx7OE(Glu-B1 locus) and Dx5(Glu-D1 locus) subunit genes are selected and multiple PCR systems for simultaneously identifying an important high-quality subunit on Glu-1 locus through one-step reaction are established by optimizing a PCR reaction system and thermal circulation conditions; and in addition, specific amplified products can be effectively separated by common agarose electrophoresis. Compared with single PCR reaction, the method greatly improves the detection efficiency and has reliable identification results and reproducibility through breed and group verification. The invention provides a simple, accurate, rapid and practical method for evaluating breeding materials as well as effectively identifying and selecting high molecular weight glutenin high-quality subunits of filial generation.

The invention discloses a kiwi fruit InDel molecular marker and a screening method and application thereof. Two different cold-proof Actinidia arguta genome DNAs are used as research subjects, resequencing of four variety genes is carried out, InDel primers are designed according to resequencing data, and PCR (polymerase chain reaction) amplification, agarose electrophoresis and non-denaturing polyacrylamide gel electrophoresis are carried out; polymorphism detection is carried in Actinidia arguta to screen out 14 pairs of InDel primers, the 14 pairs of InDel primers are applicable to the genetic diversity analysis for Actinidia chinensis, Actinidia deliciosa and the like and are also applicable to the researches, such as hybrid offspring authenticity identification, genetic map construction, and molecule-assisted breeding. The kiwi fruit InDel primers according to the embodiment of the invention has good mutation stability and low detection difficulty, allow InDel insertion/loss of large fragments, and allows agarose analysis, with steps that may be simplified.

The invention relates to a rice seedling stage cold-resistant major gene and a special primer thereof. The rice cold-resistant major gene for the special primer comprises the following steps: by taking a rice genome DNA to be detected as a template, performing polymerase chain reaction (PCR) amplification by using any one labeled primer in a labeled primer RM15040, a labeled primer ZCT13, a labeled primer ZCT23 and a labeled primer RM15123, and performing gel electrophoresis assay on the PCR amplification product to obtain rice cold-resistant identification, wherein stripes with the size of 140-170bp or 230-270bp or 290-310bp or 380-400bp exist in the PCR amplification product, and the 3rd chromosome of the seedling stage rice contains cold-resistant major genes qCTS-3-1. According to the rice cold-resistant screening, the production cost can be saved, the selection efficiency is improved, the breeding cycle of rice varieties is shortened, and the errors of the detection result are reduced.

The invention relates to the field of biotechnology-assisted seed breeding, and particularly discloses a CAPS molecular marker primer for detecting CMS (cytoplasmic male sterility) restoring genes ofcapsicum annuum and application of the CAPS molecular marker primer. CAPS molecular markers are tightly linked with the CMS restoring genes of the capsicum annuum and are M1 markers. The CAPS molecular marker primer and the application have the advantages that detection can be facilitated by the CAPS molecular markers, stable and reliable results can be obtained, the CAPS molecular markers are co-dominant markers, and accordingly sterility restoring genes of capsicum annuum strains can be detected on a large scale only by means of simple conventional experiment operation such as hereditary substance extraction, PCR (polymerase chain reaction), restriction incision enzyme digestion and routine agarose electrophoresis; the selective breeding ranges of restoring line materials for the capsicum annuum can be expanded by development and application of the CAPS molecular markers, the seed breeding efficiency can be improved, seed breeding progresses can be accelerated, and the CAPS molecularmarker primer and the application have important significance in molecular marker-assisted seed breeding for CMS three-line matched seed production and restoring line materials for the capsicum annuum.

The invention first provides a Cynoglossus semilaevis gender specific microsatellite marker, and comprises a microsatellite marker located in the Z chromosome, and the nucleotide sequence is SEQ ID NO:1. The invention also provides primers designed based on the above microsatellite marker, and the sequences of the upstream primer and the downstream primer are SEQ ID NO:2 and SEQ ID NO:3 respectively. The invention screens a Cynoglossus semilaevis gender specific microsatellite marker, and the Cynoglossus semilaevis gender specific microsatellite marker is subjected to SCAR conversion. Primers marked by SCAR are designed for Cynoglossus semilaevis heredity gender identification. Female, male and superfemale individuals of Cynoglossus semilaevis can be distinguished rapidly, accurately and effectively. Because the marker has characteristics that objective straps can be amplified in females and males and the objective straps can be distinguished with agarose electrophoresis, the time for accurate identification of Cynoglossus semilaevis heredity genders is shortened obviously, the on-site batch identification of Cynoglossus semilaevis heredity genders in a culturing farm of Cynoglossus semilaevis can be carried out, the detection cost is saved, and the work efficiency and accuracy of on-site detection of Cynoglossus semilaevis heredity genders in a culturing farm are raised.

The invention discloses a method for constructing a Dolmen structure based on a DNA origami template and gold nanorods. The method comprises the following steps: firstly, preparing gold nanorods with a specific size by virtue of a seed crystal growth method; secondly, preparing specifically designed rectangular DNA origami; and finally, adding the gold nanorods according to a mole ratio of the gold nanorods to the DNA origami of 5:1, and performing cyclic gradient annealing at 45-20 DEG C to enable the gold nanorods with the specific size to be hybridized with the specifically designed rectangular DNA origami and the Dolmen structure to be constructed with the gold nanorods. The invention also provides a preparation process for preparing the gold nanorods with the specific size by virtue of the seed crystal growth method and preparing the gold nanorods modified by a nucleotide sequence shown by ssDNA1. According to the method provided by the invention, characterization is performed with the help of agarose electrophoresis and a transmission electron microscope, a gold nanorod assembly can be positioned with a scanning electron microscope together easily by use of a characteristic that the gold nanorod assembly has a special color in a dark field image of a dark field microscope, and compared with an etching means, the method provided by the invention has the advantages of being simple, convenient and reliable, having a low cost and relatively low experiment condition requirements, and the like.

The invention provides a primer, a kit and a method for rapid identification of pigeon gender. The primer shown in the SEQ ID NO:1 and SEQ ID NO:2 has the advantages of high specificity, high accuracy, high resolution and the like in design of CHD gene sequence in pigeons; the amplification result is directly carried out by agarose gel electrohoresis, which provides higher identification so that the amplification result is intuitive and clear, and the pigeon gender can be easily distinguished. The invention further provides a molecular identification method of the pigeon gender by the aid of the primer. In the method, efficient nucleic acid lysate is further provided on the basis of the above the specific amplification of the primer, the process of obtaining DNA from feather pulp samples is greatly simplified, and time and economic cost in identification of the pigeon gender are reduced. The method and the kit for identification of pigeon gender with the PCR (polymerase chain reaction)-agarose electrophoresis technology and a genotyping method are simple in operation, rapid in identification, low in cost and abroad in the market utilization prospect.

The invention discloses a capripoxvirus (CPV) Taqman-MGB (minor groove binder) probe multiple real-time fluorescence quantitative PCR (polymerase chain reaction) detection primer, a kit and a detection method. The multiple Taqman-MGB real-time fluorescence quantitative PCR detection method can be used for rapid detection of Lumpy Skin Disease Virus (LSDV), Goatpox Virus (GTPV) and Sheeppox Virus (SPPV). According to the method, on the basis of conserved domains of target sequences of the LSDV, the GTPV and the SPPV, a pair of primer and three Taqman-MGB probes are designed. The method only needs two step amplification method and simple reaction conditions for fast, efficient, specific and highly-sensitive detection of an objective target sequence, is simple in operation, does not need expensive equipment and reagents, has no technical requirement on operators, is low in detection cost and short in testing time, and can avoid cross contamination caused by agarose electrophoresis so as to improve the detection accuracy and reliability.

The invention provides a male specific DNA marker of oplegnathus punctatus, wherein the sequence of the DNA fragment on a Y chromosome is shown in SEQ ID NO: 1; the sequence of the DNA fragment on a Xchromosome is shown in SEQ ID NO: 2. The DNA fragment on the Y chromosome is 12 bp more than the fragment on the X chromosome, and the fragment is the DNA fragment peculiar to the Y chromosome. According to the invention, two homologous and different DNA fragments on the X and Y chromosomes are screened from the whole genome sequence of the oplegnathus punctatus, and a method for identifying theheredity sex of the oplegnathus punctatus is further established; and the method can be used for quickly, accurately and effectively distinguishing the heredity sex of the oplegnathus punctatus. According to the method, a specific target band amplified in a male individual cannot be amplified by a female individual, and the product can be resolved by agarose electrophoresis, so that the time for accurately identifying the heredity sex of the oplegnathus punctatus is shortened, and the method is suitable for identifying the heredity sex of the oplegnathus punctatus in a simple environment of aculture farm, and the detection time and the cost are saved. The method has important significance and application value in sex identification, seedling breeding and family selection and breeding of the oplegnathus punctatus.

The invention relates to a method for preparing an antitumor drug carrier by using a rolling circle amplification technology. An amplification primer DNA is assembled on the surface of a nano-mateiral, a circular DNA amplification template and other amplification mixtures are added and then rolling circle amplification is performed, and the prepared product is a long single-stranded DNA antitumor drug carrier obtained by extension from the surface of nano-gold. The primer DNA is extended to a microsize scale by means of room temperature amplification and can be in comparison with a non-amplified gold-DNA control group by means of agarose electrophoresis; after amplification, the DNA becomes longer, resulting in delay in the electrophoresis experiment. Besides, in an atomic force microscopy image, the amplified long-chain DNA extending from the surface of the nano-material can be intuitively observed. The nano-scale biological composite structure constructed by the rolling circle amplification technology can be used for the researches on novel antitumor drug carriers.

The invention provides PCR authenticating primer and method of oryza punctata. The nucleotide sequence of the primer is shown as the sequence table SEQ ID No. 1 and 2. The invention also provides a PCR detection method of the oryza punctata, and the detection method comprises the steps of carrying out PCR amplification by using a sample total DNA as a template through the primer and determining a result according to agaroseelectrophoresis after the reaction is ended. The primer has good specificity, and the detection method is quick and simple and has high accuracy and good sensitivity. The invention provides an effective detection method for the authentication of the oryza punctata.

The invention provides a kit capable of simultaneously detecting 12 diarrhea pathogenic bacteria and application thereof, and belongs to the technical field of multiplex PCR detection. The kit comprises specific chimeric primers corresponding to the 12 common diarrhea pathogenic bacteria and a pair of internal reference primers, and the nucleotide sequences of the specific chimeric primer pairs are shown in SEQ ID NO.1-32 respectively. According to the kit, reaction conditions are optimized by arranging a multiplex PCR system, and accurate detection on the 12 diarrhea pathogenic bacteria is achieved by analyzing the length of a product through single-tube one-time PCR and agarose electrophoresis. The kit has the advantages of being accurate in detection, high in sensitivity, strong in specificity, simple, convenient and rapid and suitable for preliminary screening of common pathogenic bacteria in clinical diarrhea samples and epidemiological investigation of diarrhea, and a good application prospect is achieved.

The invention provides a cynoglossus semilaevis sex chromosome-linked DNA segment and application thereof. A sequence of a DNA segment on a Z chromosome is SEQ ID NO:1; a sequence of a DNA segment on a W chromosome is SEQ ID NO:2. The DNA segment is used for designing a molecular marker for identifying the genetic sex of cynoglossus semilaevis. Two Z-W chromosome homologous and different DNA segments are screened from cynoglossus semilaevis whole genome information, a primer is designed for identifying the genetic sex of the cynoglossus semilaevis, and the genetic sex of the cynoglossus semilaevis can be distinguished quickly, accurately and effectively. By adopting a method, specific purpose stripes can be amplified in male and female individuals and can be distinguished by agarose electrophoresis, the time for accurately identifying the genetic sex of the cynoglossus semilaevis is shortened, the method is suitable for identifying the genetic sex of the cynoglossus semilaevis in a simple environment of a farm, and detection time and cost are reduced.

The invention provides a primer for PCR identification of kidney beans and a PCR identification method, wherein the primer can amplify chloroplast trnL genes of the kidney beans to generate specific amplified fragments of the kidney beans, and concrete nucleotide sequences are shown as sequence tables SEQ ID No. 1&2 and No. 3&4. The PCR identification method of the kidney beans comprises the following steps: utilizing the primer for PCR amplification by taking the total DNA of a sample as a template, and judging results according to agarose electrophoresis after reaction is finished. The primer has good specificity, high accuracy and good sensitivity, and a rapid and simple detection method is provided for identifying the species resource of the kidney beans.

The invention discloses a huge creatine-kinase agarose electrophoresis agent box and making method in the medical biological domain, which comprises the following parts: agarose gel plate, Tris buffer, soluble bartitone-barbitone electrophoresis buffer, substrate, substrate buffer and color-developing agent. The invention can test serum sample to avoid false positive occurrence of CK-M B due to immune inhibiting method, which provides correct testing report for clinical.

The invention discloses a gene detection kit which is used for detecting cell chimerism or individual recognition, in particular relating to an oligonucleotide array sequence, a kit and a method usedfor detecting cell chimerism or individual recognition. The kit utilizes gene insertion/deletion polymorphism (INDELs) and site-specific fluorescent quantitative PCR to quantitatively detect that different individuals of human beings have chimerism or microchimerism of foreign body cells, which can be used for the detection of mother and son/daughter cell microchimerism, cell chimerism of donor/receptor after the transplantation of hematopoietic stem cells or other organs and residual diseases after the transplantation of hematopoietic stem cells of leukemia, and for the forensic individual identification, paternity test and the like. The kit in the invention has good specificity and high detection sensitivity, can accurately and quantitatively detect with convenient operation and with norequirement of special devices; and when adopting the method in the invention to carry out individual identification detection, only agaroseelectrophoresis is needed to distinguish the results.

The invention relates to the technical field of biological gene detection, and aims to provide an ovarian premature senility related gene whole exon amplification and detection method. The method comprises the following steps: extracting genome DNA from a detection material; using totally 120 primers shown in SEQ ID NO:1-120, carrying out an amplification test on all exons of 9 genes, namely FMR1,FOXL2, FSHR, POF1B, INHA, NOBOX, GDF9, BMP15 and FIGLA, wherein totally 61 PCR reactions in the amplification test and the negative control are carried out synchronously, and the Tm value range of the 120 primers is 57-62 DEG C; detecting an amplification product by adopting agarose electrophoresis and a gel imaging system; and analyzing the sequencing result of the amplification product by usingan automatic sequencer. All exons of 9 genes related to premature ovarian failure are synchronously amplified through optimized PCR amplification specificity, and a detection raw material is providedfor downstream detection. The method is simple and convenient to operate, short in amplification time, high in specificity and good in repeatability. The detected positive rate is greatly improved, and the application prospect is wide.

The invention relates to a small-DNA-molecular-weight standard, and a standard plasmid and a preparation method thereof. Small DNA sequences with the length of 100 bp, 200 bp, 300 bp, 400 bp, 500 bp, 600 bp, 700 bp and the like are constructed onto a plasmid by using overlapping polymerase chain reaction, restriction endonuclease digestion, DNA connection and other methods. After the plasmid is completely digested by single restriction enzyme, and seven uniform-brightness strips can be obtained through enzyme digestion. The small-DNA-molecular-weight standard is used for preparing a DNA standard reference substance, and is used for indicating the relative value of DNA molecular weight in agarose electrophoresis.

The invention discloses animal chlamydia TaqMan-MGB probe multiplex real-time fluorescent quantitative PCR (polymerase chain reaction) detection primers, kit and method. The multiplex TaqMan-MGB probe real-time fluorescent PCR method can perform quick detection on Ch. Abortus, Ch.pecorum and Ch.psittaci. On the basis of the conserved region of the three chlamydia target sequences MOMP, the detection method designs three pairs of primers and three TaqMan-MHB probes. The method can quickly, efficiently, specifically and sensitively detect the target sequences by a two-step amplification process under simple reaction conditions, is simple to operate, does not need any expensive instrument or reagent, and has the advantages of no technical requirements for operating personnel, low detection cost and short detection time. The method can avoid cross contamination which can possibly occur due to agarose electrophoresis, thereby enhancing the detection accuracy and reliability.

The invention relates to a gallinaceous visfatin gene 9bp indel polymorphism detection method and application thereof. The detection method comprises the following steps of: designing a primer in the position of a gallinaceous visfatin gene 9bp indel locus; then carrying out PCR amplification; and detecting the polymorphism of the gene 9bp indel by the polyacrylamide gel electrophoresis, wherein the gene is of a DD genotype when 9bp is deleted, the gene is of an II genotype when 9bp is inserted, and the heterozygous individual at the locus is of an ID genotype. The correlation analysis combining the detection results and economic characters of chickens is used for the assistant selection and the molecular breeding of chickens. The detection method does not need be subject to the enzyme digestion, thereby saving the cost and the time; the detection is carried out by adopting the polyacrylamide gel electrophoresis, thereby overcoming the limitation of the agarose electrophoresis to the small-fragment nucleotide difference detection. The method has the advantages of high resolution, high detection sensitivity and accurate genotype judgment; and the gel has on requirements for the staining method, and the ethidium bromide (EB) staining and the silver staining are both applicable.

The invention provides a method for identifying a self-sterile S-gene type SSR molecular marker of a sweet cherry variety. The method uses PTCR4SSR to mark, wherein the SSR primer sequence is upstream primer 5'-CATAGGTTCAAACCATACCCGTG-3' and downstream primer 5'-CTCATCTTTGTAGGGTATAATACC-3' to amplify the DNA of sweet cherry of different varieties (systems), if the amplified fragment of 255bp can be amplified, pollen S-gene exists, and a electrophoretogram can be obtained by agar gel electrophoresis; according to the agarose elelctrophoretogram, the self-sterile gene type of each variety (system) can be determined, wherein the same bands are the same S-gene type, otherwise being different S-gene types. The SAM method is used to develop SSR marked PTCR4 in sweet cherry for identifying the S-gene type of different sweet cherry self-sterile gene type varieties (systems) and guiding selection of breeding parents and configuration of pollination varieties thereof, so that the detection efficiency of the self-sterile gene type can be improved, the breeding process of the sweet cherry is shortened, and the production cost is saved.

The invention belongs to the technical field of molecular biology, and provides a molecular marker Caps7 in a paddy rice cadmium accumulation related gene OsHMA3 and an application thereof. A forwardprimer and a reverse primer are synthesized, paddy rice DNA is amplified in a PCR system, and then enzyme digestion is carried out for amplification products, finally agarose electrophoresis is used for detecting and determining that the paddy rice to be measured contains HMA3-1 or HMA3-2 allele, so that paddy rice cadmium accumulation trait is determined and seed selection of low cadmium accumulation variety of indica type rice is assisted. Paddy rice cadmium accumulation traits can be effectively determined, and a new technical means is provided for selecting low cadmium accumulation varieties of indica type rice with improved seed selection efficiency.