32 results about "Molecular cytogenetics" patented technology

The present invention provides a novel, highly sensitive and specific probe panel which detects the type of renal cortical neoplasm present in a biopsy sample. As such, the invention permits diagnosis of the predominant subtypes of renal cortical neoplasms without the use of invasive methods. The present invention further provides a molecular cytogenetic method for detecting and analyzing the type of renal cortical neoplasm present in a renal biopsy sample.

The invention discloses a poplar root tip 3D fluorescence in-situ hybridization method based on paraffin sections, and belongs to the field of molecular cytogenetics. The poplar root tip 3D fluorescence in-situ hybridization method mainly includes the steps that tender root tips of poplar are obtained and fixed with Carnoy fluid; n-butanol gradient solution dehydrating, dipping wax and embedding are carried out; section slicing, section spreading, section baking and dewaxing are carried out; section preparation adopts pectinase, cellulase, pepsin and RNase for treatment; section preparation, probe denaturation and hybridization overnight are carried out; and hybridization signals are detected after incubation with secondary antibodies is carried out. The poplar root tip 3D fluorescence in-situ hybridization method can obtain clear FISH signals of the poplar root tip paraffin sections, and 3D images captured by a laser scanning confocal microscope can accurately reflect the three-dimensional level spatial distribution of a target sequence in cells. The poplar root tip 3D fluorescence in-situ hybridization method can be used in research fields such as chromosomal space occupation ofpoplar, gene expression and analysis of Hi-C sequencing results, and has broad application prospects.

The invention belongs to the field of molecular cytogenetics and particularly relates to a method for identifying D genome and D sub-genome complete chromosome. The method comprises the following step: by taking a sequence as shown in SEQ ID NO.1 as a probe to perform fluorescence in-situ hybridization on the DNA (deoxyribonucleic acid) of a cotton genome. The sequence is taken as the probe, and chromosomes in the mitosis metaphase of the upland cotton, the asiatic cotton and the Raymond's cotton are taken as target DNA to perform fluorescence in-situ hybridization, and the results indicate that obvious hybrid signals are shown on the D subgenome chromosome of the upland cotton and the D genome chromosome of the Raymond's cotton, the signals are distributed on all chromosomes, and no obvious signals are shown in the A subgenome chromosome of the upland cotton and the A genome chromosome of the asiatic cotton. The sequence adopted by the method disclosed by the invention can be used for quickly identifying the cotton D genome and D sub-genome complete chromosome by virtue of a FISH (fluorescence in situ hybridization) method.

The present invention relates to the technical fields of bioinformatics and molecular cytogenetics, and in particular to a new method for accurately counting chromosomes by using sugarcane centromere-specific repeat sequences, comprising the following steps: step 1, obtaining centromere-specific repeat sequences of the genus Sugarcane ; Step 2, separating the Saccharum centromere repeat sequence and carrying out counting analysis; the present invention utilizes the chromatin immunoprecipitation of the Saccharum centromere-specific histone and the high-throughput sequencing technology (ChIP-Seq) to obtain The DNA sequence of the centromere region of sugarcane was obtained; through comparative analysis, the repetitive sequence in the centromere region of different sugarcane species was screened out; primers were designed according to the sequence, amplified and labeled for mitotic metaphase fluorescence in situ hybridization (FISH) , for chromosome counting. The results showed that this sequence produced specific and clear and bright signals in the centromere region of all chromosomes of different sugarcane species; the number of chromosomes of different materials could be quickly and accurately obtained by counting the number of centromere signals.

The invention discloses a method for creating and identifying a chromosomal translocation line between the genomes of peanut A and B. The method for creating is to use the cultivar Silihong as the material, and use Co 60 ‑γ-ray irradiated peanut plants to obtain peanut plants with chromosomal translocation between A and B genomes. The present invention adopts florescence Co 60 ‑γ-irradiation induced a large number of translocations between genomes A and B, realizing chromatin exchange between genomes A and B, providing new resource materials for peanut breeding and molecular cytogenetic research, and also for distant hybridization Artificial creation of exogenous chromosome (segment) introgression lines in progeny materials provides a way. The present invention uses genome fluorescence in situ hybridization technology to identify chromosome translocation lines between peanut A and B genomes, creates and identifies a batch of peanut cultivars A and B genome chromosome translocation lines new materials for the first time, and provides a basis for identifying peanut chromosome translocations effective means.

The invention discloses a method for displaying centromeres and short arms of large yellow croaker, and belongs to the field of molecular cytogenetics. A molecular marker is a segment of a 28S rDNA sequence cloned from human saliva, and is processed to prepare a probe (named as H-P3K), and the probe is crossed with metaphase chromosomes of Larimichthys crocea to obtain a positive signal. The positive signal in the centromeres and the short arms of every metaphase chromosome can be detected when FISH of the Larimichthys crocea chromosomes is carried out by the probe, the short arm signal intensity of one chromosome is weaker than the centromere signal intensity of the same chromosome, and the centromere signal intensities of different chromosomes are different. The method is used for determining the position of the centromere, analyzing the chromosome karyotype and researching the chromosome structure; and the positive signal can be used as a chromosome identifying and pairing marker.

The invention belongs to the field of molecular cytogenetics and particularly relates to a BAC-FISH method for simultaneously identifying whole sets of chromosomes of subgroups under a gossypium AD / ADE background. The method comprises the following steps: carrying out BAC-FISH on different cotton seeds by using BAC cloned 57I23 originated from an Africanum BAC library, and simultaneously identifying chromosomes of an AD genome and AD sub genome of cotton obviously; by using BAC-DNA as a probe, covering most regions out of ends of all chromosomes of the subgroup A in five gossypium tetraploids as well as a small region near the middle part of the all chromosomes of the subgroup D by a hybrid signal; and in allohexaploid hybrids AADDEE hybridized by upland cotton and G.stocksii (E1) in the group E, as the hybrid signal is similar to the tetraploid, obviously differentiating the hybrid signal from the subgroup E (without a signal) so as to simultaneously identify three sets of chromosomes of subgroups (A,D,E). By adopting the BAC-FISH method through BAC cloning, the whole sets of chromosomes of subgroups under the gossypium AD / ADE background can be simultaneously identified.

The invention discloses a method for identifying all the chromosomes of poplar by using oligonucleotide probes. According to the genome information of Populus trichocarpa, the specific oligonucleotide sequence of each chromosome is obtained through bioinformatics analysis, and the synthetic poplar 1 The oligonucleotide library corresponding to chromosome 19 is then labeled with biotin or digoxin into oligonucleotide probes, and then the chromosomes of poplar are identified by fluorescence in situ hybridization technology. Each round of hybridization can be used at most at the same time The oligonucleotide probes of the three chromosomes were repeated 7 times on the same metaphase division phase, and the 19 pairs of chromosomes of Populus species could be accurately identified. It has the advantages of common use in Populus species, good experimental repeatability, and high resolution; the invention provides a new method for accurately identifying each chromosome of Populus species, and has laid a solid foundation for the molecular cytogenetics research of Populus Base.

The invention discloses a 3D fluorescence in-situ hybridization method for a poplar root tip based on a frozen section, belonging to the field of molecular cytogenetics. The method comprises the following steps: acquiring a poplar root tip, fixing the poplar root tip with a Carnoy's fixative, then performing washing with water, embedding the poplar root tip with an embedding agent, carrying out freezing, conducting slicing, adhering an obtained section onto a glass slide, carrying out FISH steps such as section baking, denaturation, hybridization, antibody detection and the like, and finally carrying out observing under a laser confocal microscope and capturing a 3D image. The method provided by the invention has the advantages that the complete tissue structure of the root tip can be obtained and a FISH signal is clear. Compared with a 2D FISH result obtained by a traditional section preparation method, the method of the invention has the advantages that the method can visually display real space occupation of chromosomes in cells at a three-dimensional level and accurately locate the space distribution of target sequences in the cells at the three-dimensional level, is applicableto the fields of chromosome space occupation and gene expression of poplars, analysis of Hi-C sequencing results and the like, and has wide application prospects.

The invention discloses a probe combination and method suitable for the physical location of plant chromosome rDNA, and relates to the technical field of molecular cytogenetics. The probe combination includes the probes of SEQ ID NO.1-4, which are respectively 5S rDNA probe, 5.8S rDNA probe, 18S rDNA probe and 25S rDNA probe; the probe combination and method are used to carry out plant chromosome priming In situ hybridization, which can capture higher resolution chromosome and fluorescence in situ hybridization signals, and is suitable for chromosome in situ hybridization of various plants; and the probe combination is an oligonucleotide probe, which has the advantages of low production cost and convenient use , Simple operation and so on.

The invention belongs to the field of molecular cytogenetics, and in particular relates to a method for re-hybridizing a FISH film. The method mainly comprises the following steps: soaking the film into 2*SSC to wash away a cover plate after first fluorescence in situ hybridization (FISH), and then soaking the treated film in 2*SSC for 3*5min; putting the film into 2*SSC of 90 DEG C for 10min while shaking continuously; putting the film into 70% deionized formamide of 78 DEG C for 4min while shaking continuously; and quickly putting into 70% alcohol of -20 DEG C. A hybridization signal can be washed away by adopting the method, and then the hybridization signal cannot be detected by observing the film. When the treated film is reused for a FISH experiment by adopting the same probe, the hybridization signal can be detected again at the same position of a chromosome. By adopting the method, the film subjected to in situ hybridization can be reused, so that resources are effectively saved, the cost is reduced and the environmental pollution is reduced.

The invention belongs to the field of molecular cytogenetic, and concretely relates to a method for marking chromosome terminals of cotton A genome and A sub-genome. The method comprises: using BAC coming from gossypium barbadense pima-90BAC library to clone 350B21, performing BAC-FISH on mitosis mid-term chromosomes of different cotton species, and discovering the phenomena that the terminals of all chromosomes of A genome and A sub-genome generate strong hybridization signals, while the hybridization signals of all chromosomes of D genome and D sub-genome are not obvious. By utilizing the BAC-FISH method for BAC cloning, the chromosome terminals of cotton A genome and A sub-genome can be rapidly effectively marked.

The invention belongs to the technical field of crop molecular cytogenetics, and in particular relates to a method for fluorescent in situ hybridization of Chinese cabbage mature pollen cells. The method comprises steps of determining flower buds in a Chinese cabbage mature pollen stage, preparing a mature pollen cell sheet, conducting probe labeling, conducting in situ hybridization, conducting signal detection and the like. The method provided by the invention is characterized in that the flower buds before 1-2d of blossoming of Chinese cabbage are prepared, fresh pollen grains are separated from the flower buds in a 50% glacial acetic acid solution, and pollen walls are broken by virtue of vertical tabletting, so that complete vegetative cells and generative cells are released; cell samples, which are pre-processed, are denatured together with a denaturation probe at 80 DEG C for 3min, and the in situ hybridization is directly conducted, so that a fluorescent hybridization picture that signals are clear is obtained, showing a signal difference of target DNA in the vegetative cells and generative cells. The method provided by the invention fills a technical gap in fluorescent in situ hybridization of mature pollen cells of brassica plants; and the method is applicable to molecular cytogenetics study on pollen development of the Chinese cabbage.

The invention discloses an antigenic polypeptide of poplar functional centromere histone CENH3 and application thereof, belonging to the fields of bioinformatics and molecular cytogenetics. The invention mainly includes: screening a specific amino acid sequence of Populus trichocarpa CENH3 protein and artificially synthesizing the polypeptide, using the polypeptide to immunize New Zealand rabbits, preparing antiserum, and obtaining CENH3 protein antibody after affinity purification; the antibody is detected by immunofluorescence, ChIP-FISH Specificity verification proved that the antibody can accurately identify functional centromeres. The antibody can be used universally in poplar species, not only can accurately mark the position of poplar functional centromere, but also lay a foundation for the study of the structure, function and evolution of poplar centromere. The invention has broad application prospects in the research fields of poplar molecular cytogenetics, genomics, epigenomics and the like.

The invention belongs to the technical field of molecular cytogenetics and particularly relates to a method for separating cotton chromosomes, which comprises the following steps: preparing material, preprocessing the material, performing primary hypotonic treatment, performing enzymolysis, performing secondary hypotonic treatment, drying and covering a plate, removing the cover plate, performingchromosome microisolation and collecting the chromosomes. In the method of the invention, the primary hypotonic treatment, emzymolysis, secondary hypotonic treatment and plate covering method are combined to prepare for making a chromosome membrane sheet; cell walls are completely removed; the chromosomes well disperse after being compressed slightly; and the shapes of the chromosomes are so clear as to facilitate the recognition of target chromosomes.