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Microsatellite Loci, Primers and Applications of Yellow-tailed Pomfret

A technology of microsatellites and yellowtail catfish, which is applied in the determination/inspection of microorganisms, climate change adaptation, biochemical equipment and methods, etc., can solve the problem of weak molecular genetic basis, lack of high-quality genetic linkage maps, gene sequences and molecular markers Resource scarcity and other issues to achieve the effect of high polymorphism and stability

Active Publication Date: 2022-05-24
JIANGXI INST OF AQUATIC SCI
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Problems solved by technology

At present, the research on the breeding technology of yellow-tailed pomfret is still in its infancy, its molecular genetic foundation is weak, gene sequence and molecular marker resources are scarce, and there is a lack of high-quality genetic linkage maps that can be used for QTL mapping and molecular marker-assisted breeding, which seriously affects my country. The Development of the Genetic Breeding Program of Yellow-tailed Pomfret

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  • Microsatellite Loci, Primers and Applications of Yellow-tailed Pomfret

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Experimental program
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Embodiment Construction

[0022] 1. Genomic DNA extraction:

[0023] 1. Extract the genomic DNA of yellow-tailed catfish by phenol extraction method:

[0024] 1) Each individual weighed 100 mg of caudal fin into a 1.5 mL Eppendorf tube, and added 450 µL STE extraction buffer (10 mmol / L Tris-HCl, pH8.0; 1 mmol / L EDTA, pH8.0), 35 µL SDS ( 10%), 15 µL proteinase K (0.2%).

[0025] 2) After inversion and mixing, put the Eppendorf tube into a 55 ℃ water bath for digestion for about 3 hours until the solution is clear and transparent.

[0026] 3) Add RNaseA to the final concentration of 20 μg / ml, and incubate at 37 °C for 1 h.

[0027] 4) Add 700 µL of Tris saturated phenol, shake and mix on a DNA mixer for 30 min, centrifuge at 12,000 rpm for 10 min at 4 °C, and transfer the supernatant to another clean eppendorf tube (note the 1 Pipette the supernatant with the head of the mL tube to prevent confusion of the lower sediment).

[0028] 5) Add an equal volume of phenolform alcohol mixture to the supernata...

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Abstract

The invention discloses a yellow-tailed pomfret microsatellite site, a primer and an application thereof, and belongs to the technical field of molecular biology DNA markers. The nucleotide sequence of the microsatellite site is SEQ ID NO: 1, and the sequence of the microsatellite primer is SEQ ID NO: 9‑10. The present invention screens out 8 microsatellite loci from the genomic DNA of the yellow-tailed pomfret, and designs specific primers according to the flanking sequences of these loci at both ends of the microsatellite loci, and the amplification result of the obtained primers has a high degree of polymorphism And stability, it can be used in the fields of genetic diversity detection, individual identification and molecular assisted breeding of yellow-tailed pomfret, making it a reliable and effective molecular marker.

Description

technical field [0001] The present invention relates to the technical field of molecular biology DNA labeling, in particular to yellowtail microsatellite sites and primers. Background technique [0002] Yellowtail ( Xenocypris davidi Bleeker, commonly known as yellow tail, yellow flakes, yellow aunt, cucumber fish, etc., belongs to Cyprinidae, Xenocyprininae, Xenocypris, is a bottom fish, usually living in the middle and lower layers of rivers and lakes It especially likes to inhabit the bottom of waters with many aquatic plants and soft muddy bottoms. It is a small and medium-sized freshwater fish. It has strong adaptability and wide distribution. It is distributed in the Yangtze River, Pearl River, Minjiang River and various streams in southeastern Fujian in my country. Because of its thick meat, quality, delicious taste, high nutritional value, and water quality control, it has become one of the preferred varieties in the current aquaculture species structure adjustmen...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCY02A40/81
Inventor 张燕萍章海鑫王生李涵阙江龙
Owner JIANGXI INST OF AQUATIC SCI