A method for displaying the centromere and short arm of large yellow croaker

A centromere and large yellow croaker technology, applied in the field of molecular cytogenetics, can solve the problems of poor staining image quality, unsuitable for genomics and functional gene research, and difficult identification of chromosomes and fragments

Inactive Publication Date: 2018-11-23
JIMEI UNIV
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Problems solved by technology

[0003] There are still few reports on the chromosome karyotype of large yellow croaker, and the quality of staining images in most reports is poor; it basically stays in the traditional karyotype analysis, and the identification of chromosomes and fragments is still difficult
This status quo is no longer suitable for the current rapid development of genomics and functional gene research needs

Method used

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  • A method for displaying the centromere and short arm of large yellow croaker
  • A method for displaying the centromere and short arm of large yellow croaker
  • A method for displaying the centromere and short arm of large yellow croaker

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Embodiment

[0030] (1) Chromosome preparation

[0031] Inject 8mg of Codonopsis pilosula decoction / 20μg BSA mixture into the base of pectoral fin 20h in advance according to each gram of fish body weight, and temporarily store in 25℃ seawater; inject the same reagent by the same method 5 hours in advance; 2.5h in advance according to the weight per gram of fish Inject 0.5μg / g of colchicine into the base of the pectoral fin; take the head kidney cells, use 0.075M KCl to hypotonicize for 40 minutes, and fix with Carnot's fixative for 3 times; change the Carnot's fixative to methanol / volume ratio before dropping the tablets Acetic acid=2:1 mixture; put a layer of water vapor on the surface of the glass slide, add the cell suspension to 1 cm of the glass slide, and place it in the air to dry naturally;

[0032] (2) Extraction of human genomic DNA

[0033] Use tissue genomic DNA extraction kit (Shanghai Jierui) to extract human saliva DNA; before brushing your teeth for breakfast, pour normal saline...

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Abstract

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.

Description

Technical field [0001] The invention belongs to the technical field of molecular cytogenetics, and relates to a chromosome gene positioning method, in particular a method for displaying the centromere and short arms of large yellow croaker. Background technique [0002] Large yellow croaker (Larimichthys crocea), commonly known as "croaker" or "cucumber fish", belongs to the order Perciformes, Scieanidae, and Larimichthys. It is a warm-water cluster migratory fish, mainly Distributed in the southern part of my country's Yellow Sea, East China Sea, Taiwan Strait and South China Sea, it is a fish species unique to my country's coastal waters and one of the four major marine economic fishes. The scale of nursery breeding and the number of employees rank first among marine aquaculture fish. Large yellow croaker has 48 chromosomes and no sex chromosomes have been found. The length of the chromosomes is relatively short, the size of 24 pairs of chromosomes decreases successively, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6841
CPCC12Q1/6841C12Q2563/107C12Q2543/10
Inventor 曹款蔡明夷
Owner JIMEI UNIV
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