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Haliotis discus hannai microsatellite molecular marker and preparation method thereof

A molecular marker, a technology of wrinkled abalone, applied in the directions of biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of influence, unclear genetic background of aquaculture, decline of wild germplasm resources, etc. , to achieve good repeatability

Active Publication Date: 2013-05-15
DALIAN HAIBAO FISHERY
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  • Claims
  • Application Information

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Problems solved by technology

However, so far, there has been no relevant report on the microsatellite DNA molecular markers of Abalone rugosa, and it is impossible to investigate the genetic resources of wild Abalone and carry out breeding work. The decline of germplasm resources has been affected to a certain extent

Method used

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  • Haliotis discus hannai microsatellite molecular marker and preparation method thereof
  • Haliotis discus hannai microsatellite molecular marker and preparation method thereof
  • Haliotis discus hannai microsatellite molecular marker and preparation method thereof

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Embodiment Construction

[0039] Prepare microsatellite molecular markers of wrinkled disc abalone according to the following method

[0040] (1) Preparation of Genomic DNA of Abalone wrinkle

[0041] Cut about 100 mg of live wrinkled abalone mantle into a 1.5 ml centrifuge tube, cut the tissue into pieces with clean scissors, and add the following reagents to the centrifuge tube in sequence: 500ul STE buffer; 25ul proteinase K (10 mg / ml) ; 75ul 10% SDS, mix well and digest at 56°C overnight (mix once in a while). Then add an equal volume of saturated phenol solution, gently invert and mix for more than 5 minutes, centrifuge at 7000r / min for 5 minutes, and carefully transfer the supernatant to another clean centrifuge tube. Add an equal volume of phenol: chloroform: isoamyl alcohol (25:24:1) for extraction once, and centrifuge at 12000 r / min for 10 min. Then add an equal volume of chloroform:isoamyl alcohol (24:1) to extract once, centrifuge at 12000 r / min for 10 minutes, and take the supernatant. A...

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Abstract

The invention discloses a haliotis discus hannai microsatellite molecular marker which can analyze the genetic diversity of haliotis discus hannai wild population and identify a genetic relationship between cultured stocks and a preparation method thereof. According to the steps of establishing a haliotis discus hannai microsatellite (AC)n library, screening microsatellite sequence positive clone, designing a microsatellite primer, performing polymerase chain reaction (PCR) amplification and screening, ten highly polymorphic microsatellite molecular markers of Hd1, Hd2, Hd3, Hd4, Hd5, Hd6, Hd7, Hd8, Hd9 and Hd10 are obtained; and the polymorphic detection proves that the microsatellite molecular marker has the characteristic of high repeatability, is an effective and reliable molecular marker, can establish a haliotis discus hannai microsatellite DNA marker technology system and is used for evaluating the genetic diversity of haliotis discus hannai, analyzing the genetic structure and identifying the genetic relationship between the cultured stocks.

Description

technical field [0001] The invention relates to a molecular marker technology, in particular to a microsatellite molecular marker and a preparation method of the disc abalone wrinkled abalone which can analyze the genetic diversity of wild populations and identify the genetic relationship between cultured populations. Background technique [0002] Microsatellites, also known as Short Tandem Repeats (STRs) or Simple Sequence Repeats (SSRs), are a class of sequences consisting of several nucleotide pairs (called is the core sequence, generally 1-6bp) is a simple tandem repeat consisting of repeating units. The length of the repeat sequence is highly variable among different genotypes of the same species, but the two ends of the repeat sequence are generally highly conserved. By designing primers based on this conserved sequence, the variation in the number of repeats of the core sequence can be analyzed by PCR technology, and polymorphisms can be detected between different ge...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 刘小林李杨刘明泰李丹许淑芬李旭光张艳
Owner DALIAN HAIBAO FISHERY
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