12 results about "Molecular cytogenetics" patented technology

The invention relates to the field of molecular cytogenetics and potato ploidy breeding, in particular to a potato ploidy identification method for marking and counting chromosome centromere based on a potato CENH3 antibody and a ploidy detection kit. According to the method, the chromosome centromere is marked by specifically recognizing the antibody of the potato centromere nucleosome core histone CENH3 and combining the immunofluorescence technology, so that the accurate counting of the chromosome centromere is realized at the single cell level, and the ploidy of potatoes is judged. The method comprises the following steps: preparing a leaf cell suspension, sequentially incubating a potato CENH3 antibody and a labeled secondary antibody after immobilization, and observing and counting by a microscope after redyeing. The method has the advantages of high species specificity, high throughput and low cost, is not limited by special material taking parts of root tips or shoot tips and cell division periods, can accurately identify the uploid and aneuploid of potatoes, and provides an efficient tool for potato germplasm resource evaluation and cross breeding.

The invention relates to the technical field of bioinformatics and molecular cytogenetics, in particular to a method for counting centromeres of chromosomes based on a Nuf2 antibody and application. The method comprises the following steps: performing chromosome centromere counting on a Nuf2 antibody and a centromere specific protein Nuf2 through an immunofluorescence reaction, so that the number of chromosome centromeres can be directly observed, and the number of chromosomes can be efficiently and quickly determined. Besides, due to the fact that the specific protein Nuf2 of the centromere is highly conservative in eukaryotes, the invention further provides application of the method to identification of the plant ploidy, the method is easy and convenient to operate and low in cost, the ploidy identification efficiency is remarkably improved, and a universal technical platform is provided for genetic breeding.

The invention discloses a probe, a method for efficiently recognizing licorice chromosomes and application of the probe, and belongs to the field of bioinformatics and molecular cytogenetics. According to the invention, a single copy oligonucleotide sequence of No. 1-8 chromosomes of liquorice is screened by using bioinformatics, and a chromosome specific oligonucleotide probe pool is constructed through an artificially designed universal primer sequence tag splicing strategy. Through multiple rounds of FISH technology, the chromosomes 1-8 of liquorice can be efficiently and accurately identified. The method provided by the invention solves the problem of difficulty in accurate identification caused by tiny morphology and high similarity of the glycyrrhiza chromosome, has the advantages of high resolution, strong specificity, capability of identifying a single chromosome and the like, and provides a powerful tool for follow-up glycyrrhiza chromosome karyotype analysis, evolution research and exploration of a species formation mechanism.

The invention relates to a device that provides automatic humidification, fixation and programming of the optimum humidity and temperature conditions required to obtain the ideal chromosome spreading during the preparation of spreading preparations in cytogenetic and molecular cytogenetic studies, instead of humidification by breathing on the slide, values of whcih can be easily entered within the operational temperature (20°C - 37°C, ± 0.5°C) and humidity (25% - 70% ± 2%) ranges, has a cover, in which 10 preparations can be placed at a time and from which the preparations can be easily removed, thus enables ideal chromosome spreading in in situ and non-in situ cultures, including lymphocyte, bone marrow, amniocyte and fibroblast cultures by creating a controlled temperature and humidity environment that is immediately ready to spread a new patient sample, enables moistening the slide while spreading the chromosomes onto the slide in cytogenetic spreadings, and placing the slide in the cabinet and drying it at optimum humidity and temperature after spreading the pellet on the slide, the outer body of which is made of stainless steel, the heating and cooling of which is Peltier, that can filter the odour of unwanted chemicals, and is large enough to be placed on the countertop.

The application discloses a sub-genome-specific retrotransposon of elephant grass and application thereof, and belongs to the field of bioinformatics and molecular cytogenetics. The application mainly comprises the following steps: 2 sub-genome-specific retrotransposon sequences are screened out through bioinformatics analysis of short fragment sequences of the sub-genome of elephant grass; primers are designed according to the sequences, and the genomic DNA of elephant grass is used as a template to amplify the primers through PCR, and the target fragments are recovered through purification of PCR products; and it is verified that the 2 retrotransposon sequences produce clear and bright signals on the respective 14 chromosomes of elephant grass through probe labeling, chromosome sheet preparation and fluorescence in situ hybridization. The retrotransposon sequences can be directly used for specific identification of the sub-genome of elephant grass, more accurate information is provided for identification of the sub-genome of elephant grass, and a foundation is laid for chromosome engineering breeding of elephant grass.

The invention relates to the technical field of biology, in particular to antigen polypeptide of mandulapalka functional centromere histone CENH3 and application of the antigen polypeptide. The amino acid sequence of the antigen polypeptide is shown as SEQ ID No: 3. The polypeptide is used for conventionally immunizing New Zealand white rabbits to prepare antiserum and an anti-CENH3 protein antibody subjected to antigen affinity purification. The verification proves that the antibody is effective and reliable, and the functional centromere of the cyperus esculentus can be accurately identified. The method has a wide application prospect in the research fields of molecular cytogenetics, genomics, epimics and the like of cyperus esculentus.

The present invention relates to the technical fields of bioinformatics and molecular cytogenetics, and particularly relates to a method and application for chromosome centromere counting based on Nuf2 antibody. The present invention comprises the following steps: performing chromosome centromere counting on the Nuf2 antibody and the centromere-specific protein Nuf2 through an immunofluorescence reaction, so as to directly observe the number of chromosome centromeres, and thus efficiently and rapidly determine the number of chromosomes. In addition, in view of the high conservation of the centromere-specific protein Nuf2 in eukaryotes, the present invention also provides the application of this method in identifying the ploidy of plants. The operation is simple, the cost is low, the efficiency of ploidy identification is significantly improved, and a general technical platform is provided for genetic breeding.

The invention discloses a peanut chromosome centromere specific molecular marker and application thereof, particularly relates to the technical field of molecular cytogenetics, and aims to solve the technical problem that peanut chromosome centromere is difficult to accurately recognize in the prior art. According to the specific molecular marker of the peanut chromosome centromere, the molecular marker comprises CentAh106 or CentAh342; the nucleotide sequence of the CentAh106 is as shown in SEQ ID NO. 1, and the nucleotide sequence of the CentAh342 is as shown in SEQ ID NO. 2.

The invention discloses a pennisetum purpureum subgenome specific retrotransposon and application thereof, and belongs to the field of bioinformatics and molecular cell genetics. The method mainly comprises the following steps: screening two subgenome specific reverse transcription transposon sequences through bioinformatics analysis of subgenome short fragment sequences of target grass; designing a primer according to the sequence, taking Pennisetum purpureum genome DNA as a template, carrying out PCR amplification, and purifying a PCR product to recover a target fragment; probe labeling, chromosome flaking and fluorescence in situ hybridization verify that the two reverse transcription transposon sequences generate clear and bright signals on respective 14 chromosomes of pennisetum purpureum. The reverse transcription transposon sequence disclosed by the invention can be directly used for specifically identifying the pennisetum purpureum subgenome, provides more accurate information for identification of the pennisetum purpureum subgenome, and lays a foundation for engineering breeding of the pennisetum purpureum chromosome.

The invention relates to a hybridization probe mixed solution for radiation biological dose detection as well as a preparation method and a product thereof, and belongs to the field of molecular cell genetics. The technical problem to be solved is to provide a hybridization probe mixed solution with significantly improved chromosome distortion recognition accuracy and sensitivity, higher detection efficiency and better repeatability. According to the key point of the technical scheme, the hybridization probe mixed solution is composed of a probe and a hybridization buffer solution, the volume ratio of the probe to the hybridization buffer solution is 1: (50-1000); the hybridization buffer solution is a 40%-70% formamide solution containing 10 to 20 mM of Tris-HCl or PBS (Phosphate Buffer Solution). When the hybridization probe mixed solution is used for radiation biological dose detection, the distortion identification accuracy and sensitivity are high, the radiation dose estimation precision is high, and the operation is convenient.

The present application relates to the fluorescence marker for identifying the centromere region of the chromosome of P. wallichii and its application, belongs to the technical field of molecular cytogenetics, the fluorescence marker probe is named HS-CHTZ5, the probe belongs to the centromere region repeat sequence of P. wallichii, can be combined with the centromere of P. wallichii chromosome and show fluorescence signal, and the centromere segment of P. wallichii chromosome is identified.The fluorescence marker provided by the present application can be used as a beneficial supplement to the prior art, and through in situ hybridization, the efficient identification of the centromere region of the chromosome of P. wallichii is realized, and the present application has important significance for cytogenetic map construction, chromosome karyotype analysis and different genus and species chromosome relationship analysis.

The present invention belongs to the field of molecular cell genetics and chromosome engineering technology, and specifically relates to a simple operation method for chromosome pressing combined with in situ hybridization. The present invention further uses 4% paraformaldehyde treatment for double fixation on the basis of methanol glacial acetic acid fixation of chromosome pressing, so that the chromosome structure morphology can withstand high-temperature deformation above 90 degrees, and can provide a clearer chromosome morphological structure compared with traditional methods. At the same time, the present invention also develops a corresponding universal hybridization solution system and rinsing solution system, and there is no need to explore various complicated hybridization annealing processes and rinsing processes due to different probes. Compared with the traditional chromosome pressing combined with in situ hybridization technology, the present invention has higher chromosome structure morphology clarity, while maintaining the hybridization signal, the chromosome break position can still be clearly observed in the non-signal area, which helps to provide technical support for chromosome-related genetic analysis research.