Method for displaying centromeres and short arms of Larimichthys crocea

A centromere and large yellow croaker technology, applied in the field of molecular cytogenetics, can solve the problems of unsuitability for genomics and functional gene research, poor quality of stained images, and difficulty in identifying chromosomes and fragments

Inactive Publication Date: 2016-05-04
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 a

Method used

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  • Method for displaying centromeres and short arms of Larimichthys crocea

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Experimental program
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Embodiment

[0030] (1) Chromosome preparation

[0031] Inject 8mg Codonopsis Codonopsis juice / 20μgBSA mixture at the base of the pectoral fins 20 hours in advance at the base of the pectoral fins, and temporarily raise them in seawater at 25°C; inject the same reagent with the same method 5 hours in advance; inject the same reagent at the base of the pectoral fins at the rate of 2.5 hours in advance Inject 0.5 μg / g colchicine at the base; take the head kidney cells, use 0.075MKCl hypotonicity for 40 minutes, and fix with Carnoy’s fixative solution for 3 times; before dropping the tablets, replace the Carnoy’s fixative solution with a volume ratio of methanol / acetic acid = 2:1 mixed solution; put a layer of water vapor on the surface of the slide, drop the cell suspension at 1cm from the slide, and let it dry naturally in the air;

[0032] (2) Extraction of human genomic DNA

[0033] Use the tissue genome DNA extraction kit (Shanghai Jierui) to extract DNA from human saliva; before breakf...

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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 to a method for displaying the centromere and short arm of large yellow croaker. Background technique [0002] Large yellow croaker (Larimichthyscrocea), commonly known as "yellow croaker" and "cucumber fish", belongs to the order Perciformes, the family Scieanidae, and the genus Larimichthys. It is a migratory fish in warm and warm water clusters. In the southern part of the Yellow Sea, the East my country Sea, the Taiwan Strait and the South China Sea, it is a unique fish species in my country's coastal waters, and it is one of the four major marine economic fishes. Its nursery breeding scale and the number of employees rank first among seawater cultured fish. Large yellow croaker has 48 chromosomes and no sex chromosomes have been found. Chromosome length was short, and the size of 24 pairs of chromosomes decreas...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6841C12Q2563/107C12Q2543/10
Inventor 曹款蔡明夷
Owner JIMEI UNIV
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