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Construction method for Chlymys nobilis satellite mark

A technology of microsatellite marker and construction method, applied in the field of molecular biology DNA marker technology and application, can solve the problems of complicated steps and large workload, and achieve the effects of improving efficiency, simple operation and real and reliable results

Inactive Publication Date: 2007-09-26
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

The traditional method of constructing microsatellite markers is to use the method of library screening. Most of them need to construct a library, and then screen the library, which requires radioactive isotope or biotin labeling. The steps are cumbersome and the workload is heavy.

Method used

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  • Construction method for Chlymys nobilis satellite mark
  • Construction method for Chlymys nobilis satellite mark

Examples

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Embodiment Construction

[0037] The following examples are further illustrations of the present invention and should not be regarded as limiting the present invention.

[0038] 1. Extraction of DNA

[0039] Take 100 mg of the adductor muscle of Chlamysgui scallop, shred it, place it in 0.7 ml of extraction buffer, add proteinase K (final concentration: 0.1 mg / ml), and digest overnight at 37°C. The reactant was extracted with phenol:chloroform (1:1), precipitated with isopropanol, and then dissolved in TE. DNA samples were treated with RNase (20 μg / ml) at 37° C. for 1 hour, and then purified with phenol / chloroform extraction solution. The samples were detected by electrophoresis, and the concentration was determined and stored at -20°C until use.

[0040] 2. Restriction ligation and pre-amplification before hybridization

[0041] Digest the genomic DNA of Chlamydia grandis with Sau 3AI, digest overnight at 37°C, the total volume is 100μl: Genomic DNA 10μg, 10×buffer, 10μl, 10u of DpnII, add water to...

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Abstract

The invention discloses a constructing method of micro-satellite marked scallop, which comprises the following steps: extracting DNA; connecting adapter cut by enzyme; preaugumenting before crossing; crossing to enrich micro-satellite sequence through micro-satellite marked by biotin and SAV magnetic bead; obtaining DNA segment with enriched micro-satellite sequence; augumenting the segment; constructing the enriched library with enriched micro-satellite sequence; utilizing PCR to sieve the micro-satellite sequence; obtaining the result of DNA sequence; comparing the quality of segment with enriched micro-satellite sequence; designing the primer of micro-satellite sequence; adopting PAGE to detect and sieve the augumentation, polymorphism and stability of micro-satellite; statisticallizing genetic parameter of corresponding group.

Description

technical field [0001] The invention belongs to the field of DNA marker technology and application of molecular biology, and in particular relates to a method for constructing a marker of Chlamys nobilis microsatellites, a marine economic shellfish. technical background [0002] Chlamys nobilis is a warm-water shellfish, which is distributed in Fujian, Guangdong, Guangxi and the coastal areas of Hainan in my country. It is one of the four main cultured scallops in China. In recent years, the occurrence of diseases and adverse changes in the water environment have led to a significant reduction in the production of Chlamys farreri, threatening the stability and healthy development of its industry. Carrying out the genetic selection and breeding of Chlamys scallop is one of the fundamentals to solve this problem and ensure the stable development of its industry. The genetic variation of the population is the biological basis of the adaptability of organisms to nature (viabili...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12P19/34
Inventor 王艳红喻子牛
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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