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Larimichthys polyactis microsatellite DNA molecular marker

A technology of microsatellite markers and DNA molecules, applied in the direction of recombinant DNA technology, DNA / RNA fragments, etc., can solve the problem of limited quantity

Inactive Publication Date: 2013-01-16
SUN YAT SEN UNIV
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  • Abstract
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Problems solved by technology

However, there are only 35 microsatellite molecular markers developed so far for small yellow croaker (Chen et al., Conservation genetics2009, 10:1469-1471; Li et al., Molecular Ecology Notes 2006, 6(1):188-190; Ma et al., Genetics and Molecular Research 2011, 10(3): 1455-1460), even if 20 microsatellite markers (Conservation genetics 2009, 10:1629–1631; Genetics and Molecular Research 2012, 11(2):1469-1474), the number is also very limited, and more new microsatellite molecular markers for small yellow croaker are urgently needed

Method used

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Embodiment 1

[0092] 1. Construction of microsatellite molecular marker enrichment library of small yellow croaker:

[0093] 1.1 Genomic DNA extraction:

[0094] Genomic DNA was extracted by the phenol-chloroform method, and the specific steps were as follows: Cut 20 mg of caudal fin into a 1.5 ml centrifuge tube, add 570 μL of STE buffer (10 mmol / L Tris-Cl, pH=8.0; 100 mmol / L EDTA, pH=8.0; 100 mmol / L L NaCl) and cut up the tissue. Add 10μL 20mg / ml proteinase K and 20μL 20% SDS, digest at 55-56℃. Add 600 μL of phenol:chloroform:isoamyl alcohol (25:24:1) to extract twice, centrifuge at 13000r / min for 5min, and take the supernatant. Add an equal volume of -20°C pre-cooled absolute ethanol, mix well, centrifuge at 13000r / min for 5min, discard the supernatant, wash the precipitate twice with 70% ethanol, dry at room temperature, then add 50-100μL TE buffer (1mmol / L EDTA, 10mmol / L Tris-HCl, pH=8.0) to dissolve DNA. The DNA concentration was detected by an ultraviolet spectrophotometer, dilu...

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Abstract

The invention discloses a larimichthys polyactis microsatellite DNA molecular marker. The invention adopts the technical scheme that an enhanced library of larimichthys polyactis microsatellites (GT) 17, (GGT) 9 and (CTAT) 8 is built, larimichthys polyactis polymorphic microsatellite primers are screened, and the genetic polymorphism detection is carried out on larimichthys polyactis microsatellite loca, so that 21 larimichthys polyactis microsatellite markers with rich polymorphism are developed. The larimichthys polyactis microsatellite DNA molecular marker is applicable to researches on the population genetic structure and the genetic variation level of a larimichthys polyactis population.

Description

technical field [0001] The invention relates to a molecular marker technology, in particular to a small yellow croaker microsatellite DNA molecular marker. Background technique [0002] The small yellow croaker (Larimichthys polyactis) belongs to Perciformes, Totoabaidae, and Yellow Croaker. It is a warm-temperate bottom fish widely distributed in the Bohai Sea, the Yellow Sea and the northern part of the East China Sea. It is an important marine fishery economic species in my country. It is one of the "four major fisheries" in China. In recent years, the high-intensity fishing pressure has caused the small yellow croaker resources to be overfished. In order to protect and manage the fishery resources of small yellow croaker, it is very necessary to study the population genetic structure and genetic variation level. Microsatellite molecular markers have become the most commonly used method for genetic analysis of marine fish populations due to their advantages of high polym...

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 蒙子宁王乐林浩然
Owner SUN YAT SEN UNIV
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