47 results about "Polymorphic microsatellites" patented technology

The invention discloses microsatellite marker primers used for penaeus monodon microsatellite family identification. The microsatellite marker primers include six primer pairs in total, and the six primer pairs are PM-38, PM-69, PM-92, PM-84, PM-102 and PM-114 respectively. The invention further discloses an identification method for the penaeus monodon microsatellite family and application of the microsatellite marker primers to penaeus monodon microsatellite family identification. By the adoption of the microsatellite marker primers, a paternity test platform is set up on penaeus monodon with polymorphic microsatellite markers marked by fluorescence for the first time, and the identification accuracy reaches 99%; different families and sources of the penaeus monodon can be effectively and fast identified, and a basis is provided for breeding, reproduction matching and enhancement and releasing assessment of the penaeus monodon.

A microsatellite multiple fluorescent PCR method for E. coioides paternity testing is disclosed. The method includes (1) a step of extracting individual DNAs of E. coioides; (2) a step of screening E. coioides polymorphic microsatellite primers; (3) a step of optimizing E. coioides eightfold PCR conditions and performing amplification; and (4) a step of performing paternity testing. The method combines microsatellite marking and a multiple fluorescent PCR technique, eight polymorphic microsatellite loci are screened and an eightfold fluorescent PCR system is built. The eightfold fluorescent PCR system is verified by utilizing the E. coioides family. The method is simple and rapid to operate and low in cost, can be popularized and applied in E. coioides germplasm identification, family management and improved-variety selection, and provides a novel technical means for assessment of enhancement and releasing effects.

The invention discloses a microsatellite marker paternity identification primer and method suitable for nile tilapia, blue tilapia and a hybrid thereof, and application thereof, and belongs to the technical field of fish breeding. The method screens 13 pairs of fluorescent marker microsatellite primers for paternity identification. The identification method comprises the following steps of: constructing a complete sibling family of the tilapia, the blue tilapia and the hybrid thereof; extracting genomic DNA of parents and progeny of each family; screening polymorphic microsatellite markers; performing PCR amplification on fluorescent marker microsatellite primers; and performing multiple capillary electrophoresis typing and paternity identification. The invention establishes a microsatellite marker paternity identification method suitable for the tilapia, the blue tilapia and the hybrid thereof for the first time, the identification accuracy rate is 100%, and the identification accuracy rate can reach 99.43% in practical application. The method has high identification accuracy and simple and convenient operation, and can be used for guiding population inheritance and family pedigree management of the three tilapia in the breeding process.

The invention discloses general SSR molecular markers for cinnamomum and related-genera plants of the cinnamomum and a development method and application thereof. The nucleotide sequences of the general SSR molecular markers for the cinnamomum and the related-genera plants of the cinnamomum are shown in sequence tables SEQ ID NO.1-SEQ ID NO.8. A group of the polymorphic microsatellite markers andamplification primers thereof can be directly applied to intraspecific and interspecific structures in the cinnamomum and the related-genera plants of the cinnamomum, have good universality and practicability, and are stable and efficient. As the genetic background is not clear, the cinnamomum and the related-genera plants are limited by the EST sequence quantitative limiting, EDT-SSR molecular markers cannot be difficult to largely develop at the present stage, and the problem that the EDT-SSR molecular markers cannot be difficult to largely develop is solved through the general microsatellite molecular markers developed with the method.

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 relates to a portunus pelagicus polymorphic microsatellite molecular marker, an identification method and application. The identification method comprises the following steps: portunus pelagicus genomic DNA (Deoxyribonucleic Acid) extraction, simplified genomic library construction and high-throughput sequencing, sequence analysis and microsatellite primer designing, and microsatellite marker polymorphism evaluation. According to the identification method disclosed by the invention, 17983 microsatellite loci are identified, and 16 microsatellite markers having molecular polymorphism are screened out. The identification method disclosed by the invention has the advantages that the operation is simple and safe, the accuracy is high, a result is true and reliable, and the like,and is capable of being used for quickly and massively obtaining portunus pelagicus microsatellite markers; the technical method disclosed by the invention can be widely applied to numerous species having no reference genome because of no need to know genomic information in advance, and has a wide popularization and application potential; in addition, a candidate tool can be provided for population genetic variation evaluation, genetic relationship identification and genetic linkage map construction of portunus pelagicus by 16 pairs of polymorphic microsatellite marker primers obtained by themethod.

The invention provides a method for developing polymorphic microsatellite molecular markers on the basis of multi-sample high-throughput sequencing. The method comprises the steps as follows: firstly,high-throughput sequencing data of multiple sample genomes or transcriptomes of to-be-tested species are acquired, potential satellite sits capable of being amplified are screened, microsatellite sites with primer pairs appearing only once in all sequencing reads of the potential satellite sits capable of being amplified in single sample are screened, and markers which appear in multiple samplesand have the allele number larger than 2 in all the samples are screened; finally, those markers appearing in reference genomes or transcriptomes only once are screened by establishing a reference sequence database, and those repeated markers amplifying the same fragments are eliminated. The polymorphic microsatellite markers can be effectively screened by use of high-throughput sequencing data, non-specific and repeated markers can be removed, the length of PCR amplified products of corresponding microsatellite sites can be predicted, and the efficiency of development of microsatellite molecular markers is significantly improved.

The invention discloses a haliotis discus hannai microsatellite molecular marker which can analyze the genetic diversity of haliotis discus hannai wild population and identify a genetic relationship between cultured stocks and a preparation method thereof. According to the steps of establishing a haliotis discus hannai microsatellite (AC)n library, screening microsatellite sequence positive clone, designing a microsatellite primer, performing polymerase chain reaction (PCR) amplification and screening, ten highly polymorphic microsatellite molecular markers of Hd1, Hd2, Hd3, Hd4, Hd5, Hd6, Hd7, Hd8, Hd9 and Hd10 are obtained; and the polymorphic detection proves that the microsatellite molecular marker has the characteristic of high repeatability, is an effective and reliable molecular marker, can establish a haliotis discus hannai microsatellite DNA marker technology system and is used for evaluating the genetic diversity of haliotis discus hannai, analyzing the genetic structure and identifying the genetic relationship between the cultured stocks.

The invention relates to STR (short tandem repeats) molecular markers for Sporothrix globosa and applications of the STR molecular markers. The invention provides a microsatellite locus (STR molecular marker) combination for the Sporothrix globosa for the first time and provides a set of specific primers (SEQ ID NO:26-75) capable of efficiently amplifying Sporothrix globosa populations as well as detection conditions. The microsatellite locus combination contains 25 highly polymorphic microsatellite loci (SEQ ID NO:1-25). The set of microsatellite loci for the Sporothrix globosa not only can be applied to genetic background analysis and molecular tracing of the Sporothrix globosa, but also can be applied to molecular epidemiology, population genetics and phylogenetic analysis of the Sporothrix globosa.

The invention provides crested ibis microsatellite polymorphic sites, an identification method and specific primer sequences. 12 pairs of crested ibis microsatellite polymorphism site specific primersequences are screened, and the sequences are shown as SEQ ID NO. 1-SEQ ID NO. 24. The identification method of the crested ibis microsatellite polymorphic sites is used for screening the microsatellite sites and designing high quality primers; read data of crested ibis genome sequencing is compared to a crested ibis reference genome to identify the INDEL sites; according to the INDEL sites and microsatellite site information, the polymorphic microsatellite sites and high quality primers are screened. The 12 pairs of microsatellite primers are all polymorphic sites and have strong resolving power. Using the set of microsatellite primers can carry out individual identification and parent-child relationship identification of the crested ibis economically, rapidly and correctly, be beneficialfor family lineage reconstruction and information improvement of crested ibis population, and provide data support for the formulation of a scientific crested ibis protection program.

The invention discloses a peronia verruculata microsatellite marking and screening method. The method includes the steps that a peronia verruculata transcriptome sequencing library is built for Illumina HiseqTM2000 paired-end sequencing; based on a sequence obtained through sequencing, analysis and screening are conducted on microsatellite sites through MISA; screening is conducted again through SSRHunter1.3, and it is guaranteed that a front lateral wing and a rear lateral wing of the sequence have enough length for primer design; by means of an SSR primer, genome DNA of a peronia verruculata population is subjected to PCR amplification; amplification products are detected through polyacrylamide gel electrophoresis, and genotypes of the sites are determined according to different migration distances of the amplification products during population detection. Seven polymorphic microsatellite marks are obtained so as to obtain a polymorphic map of peronia verruculata genetic variation. The method is simple and quick, results are true and reliable, and the developed SSR marks can be used for phylogenetic study between peronia verruculata population genetics and onchidium shellfishes.

An identification method and an identification kit for genetic relationships among different species among sections of populus are disclosed. The method includes following steps of: S1) subjecting populus to genetic typing by adoption of polymorphic microsatellite DNA primer pairs, and constructing microsatellite DNA finger-prints of individuals in the species; and S2) subjecting populus to be identified to genotype amplification by adoption of the polymorphic microsatellite DNA primer pairs, and determining the genetic relationships of the populus to be identified according to the microsatellite DNA finger-prints, wherein the polymorphic microsatellite DNA primer pairs comprise a primer pair 1, a primer pair 2, a primer pair 3, a primer pair 4 and a primer pair 5. By application of the technical scheme, the microsatellite DNA finger-prints are constructed by utilization of the five pairs of polymorphic microsatellite DNA primers which are high in polymorphism and free of interaction between each two, and then genetic relationships of the populus among sections are determined through the microsatellite DNA finger-prints. The method is rapid, efficient, stable and suitable for different growth periods of the populus.

The invention discloses Moschus anhuiensis microsatellite loci, primers and application of primers, Moschus anhuiensis microsatellite loci comprise nine stable polymorphic microsatellite loci of which the sequences are respectively shown in SEQ ID NO: 1-SEQ ID NO: 9. The primers of Moschus anhuiensis microsatellite loci have sequences shown in SEQ ID NO: 10-SEQ ID NO: 27. By virtue of the primers, seven individual genotyping data show that the number of alleles is 0-4, the expected heterozygosity He is 0-0.779 and the observed heterozygosity Ho is 0-0.714. Except for the locus MaI75, other loci exhibit polymorphism and meet the requirement of the individual identification.

The invention discloses a mandarin fish paternity test kit and a microsatellite PCR identification method thereof. The kit comprises primers which amplify 20 microsatellite loci. The mandarin fish paternity test kit and the microsatellite PCR identification method thereof use the combination of microsatellite markers and a PCR technology to screen the 20 highly polymorphic microsatellite loci, andconduct high-throughput individual recognition and parent-child relationship analysis of the mandarin fish family. A single PCR reaction can detect 4 or 5 sites, relatively simple unit point detection is performed, the efficiency is increased by more than 4 times, and the cost is reduced to about one-third of the original. The error in the interpretation of the intermediate gene size by polyacrylamide gel electrophoresis is overcome by parting through a sequenator, the accuracy of genotype data is improved, and the analysis result shows that 99% of the individuals to be tested can find male and female parents correctly. The establishment of the mandarin fish paternity test kit and the microsatellite PCR identification method thereof provides a new technical means for the mandarin fish germplasm identification, family management and evaluation of the effects of multiplication and emission.

The invention discloses a larimichthys polyactis microsatellite DNA molecular marker. The invention adopts the technical scheme that an enhanced library of larimichthys polyactis microsatellites (GT) 17, (GGT) 9 and (CTAT) 8 is built, larimichthys polyactis polymorphic microsatellite primers are screened, and the genetic polymorphism detection is carried out on larimichthys polyactis microsatellite loca, so that 21 larimichthys polyactis microsatellite markers with rich polymorphism are developed. The larimichthys polyactis microsatellite DNA molecular marker is applicable to researches on the population genetic structure and the genetic variation level of a larimichthys polyactis population.

The invention discloses a radix polygonati officinalis microsatellite DNA molecular marker, comprising the steps of constructing a radix polygonati officinalis microsatellite (CT) n enrichment library; screening and sequencing microsatellite sequence positive clone; obtaining 55 clones containing the microsatellite repeated sequence; and screening to obtain 9 high polymorphic microsatellite molecular markers Po1, Po2, Po3, Po4, Po5, Po6, Po7, Po8 and Po9. The radix polygonati officinalis microsatellite DNA molecular marker can be used for researching genetic diversity of the radix polygonati officinalis, variation and evolutionary relationship between polygonatum plants and population, has good repeatability and is a reliable and efficient molecular marker.

The invention discloses microsatellite marker primers used for pinctada fucata martensii microsatellite family identification. The microsatellite marker primers include nine primer pairs in total, and the nine primer pairs are PF-3, PF-4, PF-11, PF-13, PF-16, PF-27, PF-30, PF-44 and PF-48 respectively. The invention further discloses an identification method for the pinctada fucata martensii microsatellite family and application of the microsatellite marker primers to pinctada fucata martensii microsatellite family identification. By the adoption of the microsatellite marker primers, a paternity test platform is set up on pinctada fucata martensii with polymorphic microsatellite markers marked by fluorescence for the first time, and the identification accuracy reaches 90.24%; different families and sources of the pinctada fucata martensii can be effectively and fast identified, and a basis is provided for breeding, reproduction matching and enhancement and releasing assessment of the pinctada fucata martensii.

The invention discloses dasyatis zugei microsatellite sites, primers and application thereof. The dasyatis zugei microsatellite sites include 10 stable polymorphic microsatellite sites, wherein the sequences of the 10 stable polymorphic microsatellite sites are respectively as shown in SEQ ID NO:1-SEQ ID NO:10. The sequences of the primers of the dasyatis zugei microsatellite sites are respectively as shown in SEQ ID NO:11-SEQ ID NO:30. The application of the dasyatis zugei microsatellite sites or the primers comprises molecular ecology study, pedigree analysis, germplasm resource survey, assistant breeding and individual identification. The genotyping result of 10 pairs of primers in 15 dasyatis zugei individuals shows that the allelic gene number is 3-24, and the average allelic gene number is 13.6; the expected heterozygosity He is 0.5-0.984, and the observed heterozygosity Ho is 0.429-1, so that the 10 pairs of primers are indicated to have high polymorphism and meet the requirement for individual identification of dasyatis zugei.

The invention discloses a microsatellite-based paramisgurnus dabryanus family identification method and application thereof. The microsatellite-based paramisgurnus dabryanus family identification method comprises the following steps: acquiring paramisgurnus dabryanus full-sib families; extracting genomic DNA from parents and offsprings; screening polymorphic microsatellite markers, and synthesizing primers; performing fluorescence PCR amplification on DNA samples of the parents and offsprings; performing genotypic analysis on the parents and offsprings; and the like. The number of selected microsatellite site alleles is large, the polymorphism is high, and a PCR product is stable and reliable. When the microsatellite sites are used for identifying the 17 full-sib families, the probabilities of finding true male and female parents for 340 offspring individuals are 95.9% and 97.9% respectively, and the overall success rate is as high as 95.6%; by the microsatellite-based paramisgurnus dabryanus family identification method, requirements on identification of a parental sibship and family management in genetic paramisgurnus dabryanus breeding can be met, and a strong technical supportis provided for molecular breeding of paramisgurnus dabryanus.

The invention discloses a sparus latus microsatellite family identification method. The sparus latus microsatellite family identification method comprises the following steps: (1) sparus latus full-sib family establishment; (2) extracting genomic DNA (deoxyribonucleic acid) of parents and offspring of sparus latus; (3) screening polymorphic microsatellite primers; (4) developing and amplifying a fluorescence labeling 8-PCR system; (5) microsatellite site genotyping and family identification: (5.1) determining corresponding genotypes of parents and family individuals on each microsatellite site; and (5.2) performing family identification. According to the method, a paternity test platform is established on sparus latus by utilizing the fluorescence-modified polymorphic microsatellite marker for the first time, and the method is high in identification accuracy, rapid and low in cost. The invention also discloses application of the method in population genetics evaluation, genealogy authentication, paternity test, molecular marker-assisted family management and molecular marker-assisted parent selection of sparus latus.

The invention discloses a microsatellite marker paternity identification primer and method suitable for nile tilapia, blue tilapia and a hybrid thereof, and application thereof, and belongs to the technical field of fish breeding. The method screens 13 pairs of fluorescent marker microsatellite primers for paternity identification. The identification method comprises the following steps of: constructing a complete sibling family of the tilapia, the blue tilapia and the hybrid thereof; extracting genomic DNA of parents and progeny of each family; screening polymorphic microsatellite markers; performing PCR amplification on fluorescent marker microsatellite primers; and performing multiple capillary electrophoresis typing and paternity identification. The invention establishes a microsatellite marker paternity identification method suitable for the tilapia, the blue tilapia and the hybrid thereof for the first time, the identification accuracy rate is 100%, and the identification accuracy rate can reach 99.43% in practical application. The method has high identification accuracy and simple and convenient operation, and can be used for guiding population inheritance and family pedigree management of the three tilapia in the breeding process.

The invention discloses spodoptera frugiperda polymorphic microsatellite molecular markers and primers thereof. Nucleotide sequences of the microsatellite molecular markers are correspondingly shown as SEQ ID NO.1-12, and primer pairs of the microsatellite molecular marker are shown as SEQ ID NO.13-36. The invention further discloses a preparation method of the polymorphic microsatellite molecularmarker. According to the invention, polymorphism experimental verification is carried out on 33 sites by predicting four-base microsatellite sites with polymorphism in the genome of spodoptera frugiperda, and twelve four-base microsatellite markers with good polymorphism are obtained through screening for the first time, wherein the four-base microsatellite markers comprise four moderate polymorphism markers and eight high polymorphism markers, so that molecular technical support is provided for genetic research on invasion of spodoptera frugiperda, and molecular ecological research on the highly-multiphagous species, namely the grassland spodoptera litura, is facilitated.

The invention discloses a method for screening microsatellite markers of seagrass halophila. Firstly, the improved CTAB method is used to extract genomic DNA of the leaf tissue of halophila; , and the DNA fragments were sequenced, and DNA sequences with more than 5 base repeats (2-6) units were screened; then specific primers were designed at the flanking sequences at both ends of the microsatellite repeat sequence, and different geographical distribution groups The individual genome DNA of Salina philodendron was amplified by PCR to detect and optimize primers; finally, the fragment size of the PCR product was analyzed to determine the genotype of each individual, obtain the genetic polymorphism map of Salina philodendron, and determine the repeatability, Microsatellite markers with high stability and polymorphism. The invention can be used for the identification of germplasm resources, genetic diversity analysis, genetic map construction and other researches of the halophobia, and can be further used for the management, protection and restoration of the halophobia seagrass bed.

The invention belongs to the technical field of molecular biology DNA molecular markers, and discloses a polymorphic microsatellite molecular marker, primers and applications of the polymorphic microsatellite molecular marker of the balsamina chinensis. / R, 57324F / R, 29968F / R, 6321F / R, 17156F / R, 16775F / R, 23957F / R, 11365F / R, 50660F / R, 2904F / R, 45390F / R, 43219F / R, 20549F / R , 9153F / R, 31370F / R and 852F / R, the nucleotide sequences of the polymorphic microsatellite molecular markers of the fish lacking beetles are shown in SEQ.ID.No.1-12 respectively. The present invention can be used for the analysis of population genetic diversity and population genetic structure of A. chinensis.

The invention provides crested ibis microsatellite polymorphic sites, an identification method and specific primer sequences. 12 pairs of crested ibis microsatellite polymorphism site specific primersequences are screened, and the sequences are shown as SEQ ID NO. 1-SEQ ID NO. 24. The identification method of the crested ibis microsatellite polymorphic sites is used for screening the microsatellite sites and designing high quality primers; read data of crested ibis genome sequencing is compared to a crested ibis reference genome to identify the INDEL sites; according to the INDEL sites and microsatellite site information, the polymorphic microsatellite sites and high quality primers are screened. The 12 pairs of microsatellite primers are all polymorphic sites and have strong resolving power. Using the set of microsatellite primers can carry out individual identification and parent-child relationship identification of the crested ibis economically, rapidly and correctly, be beneficialfor family lineage reconstruction and information improvement of crested ibis population, and provide data support for the formulation of a scientific crested ibis protection program.

The invention discloses Hyla tsinlingensis microsatellite sites, primers for the microsatellite sites and application of the microsatellite sites and the primers. The Hyla tsinlingensis microsatellite sites include 12 stable polymorphic microsatellite sites of which the sequences are respectively shown in SEQ ID NO:1-SEQ ID NO:12. By using the Hyla tsinlingensis microsatellite sites and through analyzing genetic typing results of 18 Hyla tsinlingensis individuals, the number of alleles is 3-12, expected heterozygosity He is 0.145-0.903, observed heterozygosity Ho is 0-0.579. All the sites show relatively high polymorphism and the requirement for individual identification is reached.

The invention discloses microsatellite marker primers used for penaeus monodon microsatellite family identification. The microsatellite marker primers include six primer pairs in total, and the six primer pairs are PM-38, PM-69, PM-92, PM-84, PM-102 and PM-114 respectively. The invention further discloses an identification method for the penaeus monodon microsatellite family and application of the microsatellite marker primers to penaeus monodon microsatellite family identification. By the adoption of the microsatellite marker primers, a paternity test platform is set up on penaeus monodon with polymorphic microsatellite markers marked by fluorescence for the first time, and the identification accuracy reaches 99%; different families and sources of the penaeus monodon can be effectively and fast identified, and a basis is provided for breeding, reproduction matching and enhancement and releasing assessment of the penaeus monodon.

A microsatellite multiple fluorescent PCR method for E. coioides paternity testing is disclosed. The method includes (1) a step of extracting individual DNAs of E. coioides; (2) a step of screening E. coioides polymorphic microsatellite primers; (3) a step of optimizing E. coioides eightfold PCR conditions and performing amplification; and (4) a step of performing paternity testing. The method combines microsatellite marking and a multiple fluorescent PCR technique, eight polymorphic microsatellite loci are screened and an eightfold fluorescent PCR system is built. The eightfold fluorescent PCR system is verified by utilizing the E. coioides family. The method is simple and rapid to operate and low in cost, can be popularized and applied in E. coioides germplasm identification, family management and improved-variety selection, and provides a novel technical means for assessment of enhancement and releasing effects.

An identification method and an identification kit for genetic relationships among different species among sections of populus are disclosed. The method includes following steps of: S1) subjecting populus to genetic typing by adoption of polymorphic microsatellite DNA primer pairs, and constructing microsatellite DNA finger-prints of individuals in the species; and S2) subjecting populus to be identified to genotype amplification by adoption of the polymorphic microsatellite DNA primer pairs, and determining the genetic relationships of the populus to be identified according to the microsatellite DNA finger-prints, wherein the polymorphic microsatellite DNA primer pairs comprise a primer pair 1, a primer pair 2, a primer pair 3, a primer pair 4 and a primer pair 5. By application of the technical scheme, the microsatellite DNA finger-prints are constructed by utilization of the five pairs of polymorphic microsatellite DNA primers which are high in polymorphism and free of interaction between each two, and then genetic relationships of the populus among sections are determined through the microsatellite DNA finger-prints. The method is rapid, efficient, stable and suitable for different growth periods of the populus.