84results about How to "Polymorphism rich" patented technology

The invention relates to the field of molecular markers, and provides a method for quickly identifying purity of a watermelon variety Zhentian 1217 by using an EST-SSR (expressed sequence tag-simple sequence repeat) molecular marker. According to the method, a specially designed EST-SSR primer is adopted, a genome DNA of the Zhentian 1217 is used as a template to perform amplification, and a specific band of an amplification result is used for detecting the variety purity of the Zhentian 1217 to be detected. By adopting the method, the identification can be performed at any period of the growth of a Zhentian 1217 plant, and hybrid seeds can be differentiated from female parent selfed seeds and male parent selfed seeds; and the method is high in accuracy and broad in application popularization prospect, and provides a scientific basis for carrying out watermelon seed preparation and selling high-quality seeds timely.

The invention discloses universal primers for identifying bacillus. The universal primers comprise a forward degenerate primer composed of a nucleotide sequence shown as SEQID NO:1 and a reverse degenerate primer composed of a nucleotide sequence shown as SEQID NO:2. The invention also discloses a method for classifying and identifying bacillus, which comprises the following steps: utilizing the universal primers to perform PCR (Polymerase Chain Reaction) amplification; sequencing the PCR amplification product; utilizing sequence comparison software to perform sequence comparison in a Genbank data base; and acquiring the variety of a known strain having the highest sequence similarity, namely the variety of the strain. All varieties of bacillus can be classified by the universal primers, and the classification result is accurate and reliable; and the universal primers can be used for classifying bacillus on a variety level and can be also used for classifying bacillus on a subvariety level. Besides, the classification and identification method is simple and has a high identification speed.

The invention discloses an EST-SSR (expressed sequence tag-simple sequence repeat) marker primer group developed based on Paphiopedilum henryanum transcriptome sequences; the group includes 34 pairs of primers having nucleotide sequences shown as in SEQ ID NO. 1 to SEQ ID NO. 68. The invention also discloses an agent comprising the Paphiopedilum henryanum EST-SSR marker primers. The invention further discloses a method of developing the Paphiopedilum henryanum EST-SSR marker primers. The Paphiopedilum henryanum EST-SSR marker primers provided herein have the advantages such as rich polymorphism, good repeatability, good amplification stability and good static convenience, and are applicable to genetic diversity analysis and molecular mark assisted breeding of germplasm resources of Paphiopedilum henryanum and other species in Paphiopedilum, especially subgenus Paphiopedilum, as well as to molecular research relating to plants in Paphiopedilum.

The invention provides a method for screening microsatellite markers of Macrobrachium nipponensis by a magnetic beads enrichment method, including the following steps: the extraction of the genomic DNA of the Macrobrachium nipponensis; the hybridization of a probe and a target segment; the enrichment of magnetic beads; positive sequencing clone and the amplification of the genomic DNA according to primers which are related to the flanking sequences of the microsatellite. As the invention combines the magnetic beads enrichment and PRC amplification of bacterial liquid to carry out second-time screening, no isotope is needed. The method has the advantages of safety and high efficiency and low cost, short cycle, high reliability and the like. At the same time, in the enrichment process, the magnetic beads are repeatedly washed for a plurality of times, which can clean and remove microsatellite sequences which are not firmly adhered because of less repetition times. Therefore, the obtained microsatellite sequences are longer and are repeated for more times. The microsatellite sequences which are more frequently repeated are considered to have rich diversity at interspecies and intraspecies and can be widely applied to the diversity identification of germ plasm, the building of genetic maps, and QTL. Therefore, the method is one which screens the microsatellite markers of Macrobrachium nipponensis and can be easily popularized.

The invention discloses an InDel molecular marker interlocked with a gossypium harknessii male sterile recovery gene and application. The molecular marker is 5'-TAGAAGACTGGA-3', and is located at a position, which is distant from an initiation codon by 1247 bp in a promoter region of the recovery series gene Gh_D05G3392, and an InDel molecular marker primer is as shown in SEQID NO. 1-2. By the InDel molecular marker primer, whether recovery genes in materials exist or not and are homozygous or not is effectively identified by operations such as simple PCR and polyacrylamide gel electrophoresis, therefore, breeding of a field cotton cytoplasmic male sterile new recovery series material is accelerated, and seed purity of cotton recovery series is ensured.

The invention discloses a method for monitoring Siniperca chuatsi germplasms by virtue of microsatellite genetic marker. The method comprises genome DNA extraction and detection, microsatellite sequence search and analysis, polymorphic microsatellite primer selection and optimization, polymorphic microsatellite detection and monitoring of different geographic siniperca chuatsi populations and hereditary features thereof. Microsatellite loci selection and polymorphic microsatellite marker development and verification are performed on a large number of EST generated by siniperca chuatsi transcriptome sequencing, so that 15 EST-SSR polymorphic microsatellite markers are developed, and different geographic siniperca chuatsi populations and hereditary features thereof can be monitored; the limitations of fuzzy genetic background and less molecular marker amount in existing germplasm detection, selective breeding and germplasm improvement research of siniperca chuatsi can be broken, and a basis is provided for germplasm detection, selective breeding and germplasm improvement of siniperca chuatsi, so that germplasm detection, auxiliary genetic breeding and genomic research processes of siniperca chuatsi molecular markers are promoted.

The invention discloses an SSR (Simple Sequence repeats) primer group based on Fraxinus Velutina Torr. transcriptome sequencing information development and an application of the primer group in germplasm identification, belonging to the technical field of molecular biology. The SSR primer group comprises 42 pairs of primers, wherein the nucleotide sequences of the first pair to the forty-second pair of primers are shown in SEQ ID No.1 to SEQ ID No.84. Experiments verify that the SSR primer group disclosed by the invention has the advantages of rich polymorphism, good repeatability, clear electrophoretic band and the like. The SSR primer group can be effectively used for researches on germplasm identification, DNA fingerprint establishment and the like. A germplasm identification method is sensitive and reliable, can be used for rapidly and accurately realizing and finishing distinguishment and identification of Fraxinus chinensis germplasm, and has important significances in identification and intellectual property protection of Fraxinus chinensis germplasm resources and genetic breeding of genetic breeding molecules.

Belonging to the field of biotechnologies, the invention provides a molecular marking method for identifying a tea clonal variety of tea trees. The method includes the following steps of: 1) putting 1.0g of fresh and tender buds and leaves of a variety to be tested into a mortar, adding liquid nitrogen and grinding the mixture into powder, adding a CTAB buffer solution, conducting water bathing at a temperature of 65DEG C for 30-60 minutes, then adding a DNA extracting solution to obtain the genome DNA of the tea tree variety to be tested; 2) taking the genome DNA extracted in step 1) as a template, using SSR (simple sequence repeat) primers for PCR amplification, and subjecting the amplification product to 10% native polyacrylamide gel electrophoresis, thus obtaining an electropherogram of the variety to be tested; and 3) comparing the electropherogram of the variety to be tested with a standard tea tree variety electropherogram obtained according to the step 1) and step 2), and if the variety to be tested and the standard variety are consistent in band composition, determining the variety to be tested and the standard variety as an identical variety, thus obtaining the purity of the variety to be tested. Compared with other methods, the invention can reflect the genetic background and genetic relationship of tea tree varieties more truly. And the technology provided in the invention can be used as a scientific basis for identifying the authenticity of the tea tree varieties.

The invention relates to the technical field of molecular markers, in particular to a chenopodium quinoa willd EST-SSR (Expressed Sequence Tag-Simple Sequence Repeat) molecular marker, a development method of the chenopodium quinoa willd EST-SSR molecular marker and application of the chenopodium quinoa willd EST-SSR molecular marker. A group of chenopodium quinoa willd EST-SSR molecular markers comprises forward primers and reverse primers corresponding to 66 sites. The development method of the chenopodium quinoa willd EST-SSR molecular marker comprises the following steps of: 1, obtaining RNA-seq and EST sequences from an NCBI (National Center of Biotechnology Information) database; 2, using bioinformatics software to process, assemble and splice sequences into unigenes, and performing SSR site searching on the unigenes to obtain the EST-SSR; and 3, designing EST-SSR primer amplification chenopodium quinoa willd genome DNA for verifying the EST-SSR, and using the verified EST-SSR molecular marker to realize application of genetic diversity analysis in chenopodiaceae germplasm. The EST-SSR molecular marker developed on the basis of a public database has the advantages of high efficiency, high success rate, rich polymorphism and high universality.

The invention discloses a SSR core primer set based on Pennisetum purpureum Schum transcriptome sequence development and its application. The SSR core primer set comprises 50 pairs of primers and the primers have nucleotide sequences shown in SEQUENCE ID NO. 1 to 100 in the sequence table. The invention also relates to a use of the SSR core primer set in Pennisetum purpureum Schum genetic diversity analysis and Pennisetum purpureum Schum germplasm genetic genealogy analysis. The 50 pairs of the SSR primers are derived from Pennisetum purpureum Schum spire transcriptome sequences. Compared with the existing Chinese pennisetum common SSR primers, the SSR core primer set has the advantages of high polymorphism, stable amplification, good repeatability and statistics convenience. The primers are successfully applied to Pennisetum purpureum Schum genetic diversity analysis and Pennisetum purpureum Schum germplasm genetic genealogy analysis, increase the amount of SSR primers used by Pennisetum purpureum Schum, can be effectively used for Pennisetum purpureum Schum genetic diversity analysis, identification of variety, fingerprinting construction and molecular marker-assisted breeding, and other Chinese pennisetum related sturdy. The SSR core primer set is suitable for promotion and application in the field of biotechnology.

The invention relates to a rapid identification method for truth of corn varieties. In the method, an embryo of a single dry seed is adopted to extract DNA of a genome of a sample to be detected, and the method is time-saving and ensures that the extracted genome has high and steady quality; 20 pairs of SSR molecular marker core primers are used, have rich polymorphism and steady banding patterns, and are evenly distributed on 10 chromosomes; and the method also adopts an optimized TouchDown PCR amplification program. The rapid identification method for the truth of the corn varieties has low design cost for the primers, is simple in operation and time-saving, can conveniently and quickly identify the authenticity of the corn varieties, and is particularly suitable for material identification of a large amount of samples.

The invention discloses a Sacalia bealei microsatellite marked specific primer and a detection method. The detection method comprises the following steps: performing polymerase chain reaction (PCR) amplification on microsatellite site sequence by taking Sacalia bealei genome DNA as a template and using the microsatellite marked specific primer; performing non-denaturing polyacrylamide gel electrophoresis detection on the PCR amplification products; and performing polymorphic analysis on microsatellite sites according to the detection result. The Sacalia bealei microsatellite marked specific primer and the detection method thereof, provided by the invention, can be applied in the fields of genetic diversity level evaluation, population genetic structure analysis and relationship identification of Sacalia bealei.

The invention relates to a SNP marker used for traceability in a pig SFTPC gene and a detection method thereof. The SNP molecular marker is obtained through DNA pool sequencing, and has a sequence as shown in SEQ ID NO 1; a base is mutated from 639A to 639T at the 639th base of the SNP molecular marker, and the coded amino acid sequence is shown in SEQ ID NO 2. In a test group including 11 pig varieties or strains, the allele distribution condition of the SNP molecular marker in the test group is analyzed, and the allele frequencies of the molecular marker in different varieties or strains are found to be approximate; the polymorphism is abundant; the allele frequency distribution difference between varieties or strains is fewer; the heterozygosity is more than 0.3; and the SNP marker is preliminary determined to be applicable to DNA traceablility for pork products.

Relating to the field of plant protection, the invention specifically discloses a primer set of a puccinia recondita tritici EST-SSR molecular marker, a detection method and application thereof. The primer set of the Puccinia recondita tritici EST-SSR molecular marker includes primers shown as nucleotide sequences SEQ ID No:1-37. The detection method for puccinia recondita tritici genetic diversity employs the primer set to analyze the puccinia recondita tritici diversity and the population genetic structure, has the characteristics of simple operation, convenient detection, stable PCR amplification result, clear electrophoretic band and rich polymorphism, plays an important role in puccinia recondita tritici toxicity monitoring and genetic diversity analysis.

The invention belongs to the technical field of buchloe dactyloides molecular breeding, and particularly relates to an optimal buchloe dactyloides ISSR-PCR (inter-simple sequence repeat-polymerase chain reaction) reaction system. The invention provides the following ISSR-PCR reaction system that: each 25muL of reaction system contains 1*PCR buffer solution, 0.3mM of dNTPs, 1.0U of Taq enzyme, 0.6muM of any one of primers shown as SEQ ID 1-7, 2.0mM of Mg2+ and 50ng of DNA template; and the reaction mixed solution is amplified by the following procedures of: pre-denaturing for 3 minutes at 94 DEG C, denaturing for 30 seconds at 94 DEG C, annealing for 30 seconds at 50 to 60 DEG C, stretching for 2 minutes at 72 DEG C, circulating for 35 times, and stretching for 4 minutes at 72 DEG C. The buchloe dactyloides ISSR-PCR provided by the invention has good stability and high definition, and increases the richness of strips.

The invention discloses a primer set for hybrid cymbidium tortisepalum EST (expressed sequence tags)-SSR (simple sequence repeats) labeling. The primer set includes 4 pairs of primers, and the nucleotide sequences of the primers are as shown in the sequence tables SED ID NO.1-SED ID NO.8. The invention further discloses a screening method of the primer set for hybrid cymbidium tortisepalum EST-SSRlabeling. The primer set for hybrid cymbidium tortisepalum EST-SSR labeling obtained by the screening method has the advantages of rich polymorphisms, stable amplification and convenient statistics,and can be used for genetic diversity analysis of germplasm resources of hybrid cymbidium tortisepalum plant varieties, molecular marker-assisted breeding and related molecular research.

The invention discloses an EST-SSR (expressed sequence tag-simple sequence repeat) marker primer developed based on a platycladus orientalis transcriptome sequence and application of the EST According to the EST-SSR marker primer, an SSR sequence is obtained on the basis of platycladus orientalis transcriptome sequencing data, a primer is designed, and the stably-amplified, efficient and polymorphic microsatellite marker primer is obtained through a triple screening method of agarose gel electrophoresis, polyacrylamide gel electrophoresis and capillary electrophoresis screening. The effectiveness of the developed EST-SSR marker primer in the aspects of genetic diversity analysis and genetic relationship identification is verified by utilizing different varieties and clone of platycladus orientalis, and the EST-SSR marker primer can be applied to genetic diversity analysis, genetic relationship and variety identification, molecular marker-assisted selective breeding, genetic relationship identification and genetic relationship identification of platycladus orientalis plants. And the research fields of new plant variety protection and the like.

The invention discloses a chicken low-density SNP liquid phase chip based on targeted capture sequencing and application of the chicken low-density SNP liquid phase chip, the chip contains 5912 SNP loci, 2582 SNP loci obviously associated with economic traits of slaughter type chicken breeds are derived from whole genome re-sequencing data of 33 representative local chicken breeds in China and related researches, and the SNP loci have the advantages that the SNP loci are high in specificity and high in specificity; the method is suitable for resource evaluation and genetic identification of local chicken varieties and application to economic character genetic improvement of slaughter type broiler chicken varieties. Meanwhile, according to the chicken low-density liquid-phase chip, the target SNP sites can be adjusted by directly increasing or decreasing the probes, and compared with a solid-phase chip, the chicken low-density liquid-phase chip has better flexibility.

The invention relates to an SNP molecular maker in a pig PSMA1 gene for traceability and a detection method thereof. The SNP molecular maker is obtained by DNA pooling and sequencing and is represented by SEQ ID NO.3, and an amino acid sequence encoded by the SNP molecular maker is represented by SEQ ID NO.4. The SNP molecular maker is in a DNA fragment of the pig PSMA1 gene, wherein, the DNA fragment of the pig PSMA1 gene is represented by SEQ ID NO.1, and an amino acid sequence encoded by the DNA fragment of the pig PSMA1 gene is represented by SEQ ID No.2. The analysis of the allelic distribution of the SNP molecular maker in a group of trials containing 9 pig species/strains shows that allelic frequencies of the SNP molecular maker in different species or strains are approximate, the polymorphism is high, allelic frequency distribution difference between species or strains is small, and the heterozygosis degree is all larger than 0.3, thus the SNP molecular maker disclosed herein can be used for DNA technology for pork traceability.

The invention discloses an SSR primer group for analyzing a Hevea brasiliensis (Willd. ex A. Juss.) Muell. Arg. powdery mildew colony genetic structure, and application of the SSR primer group. The SSR primer group for analyzing the Hevea brasiliensis (Willd. ex A. Juss.) Muell. Arg. powdery mildew colony genetic structure comprises 16 pairs of primers, wherein the nucleotide sequences of the primers are as shown in SEQ ID NO.17-48. The invention also provides application of the primer group in the Hevea brasiliensis (Willd. ex A. Juss.) Muell. Arg. powdery mildew colony genetic structure analysis. The primer group has the characteristics of being good in specificity, stable in amplification, rich in polymorphism, good in repeatability, convenient in counting and the like, and a new technology and a new means are provided for research on the Hevea brasiliensis (Willd. ex A. Juss.) Muell. Arg. powdery mildew colony genetic structure.