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Method for analyzing genetic diversity of shellfish by using fAFLP (Fluorescent Amplified Fragment Length Polymorphism) labeling technology

A technology of genetic diversity and marker technology, applied in the field of analysis of shellfish genetic diversity using fAFLP marker technology, can solve the problems of enzyme digestion, pre-amplification, selective amplification without providing a reaction system, etc., to achieve gene diversity and The effect of large Shannon information index, high proportion of polymorphic sites, and many amplification sites

Inactive Publication Date: 2011-05-04
ZHEJIANG WANLI UNIV
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Problems solved by technology

At present, in terms of marine shellfish, Peng Yongxing et al. (2008) used fAFLP technology to study four species of clams (Meretrix meretrix), green clams (Cyclinasinensis), Philippine clams (Ruditapes philippinarum) and hard-shelled clams (Mercenaria mercenaria). The population genetic diversity and inter-species relationship were studied. In this literature, enzyme digestion, pre-amplification, and selective amplification were tested, and a set of reaction systems was provided for us; Lin Zhihua et al. (2009) used fAFLP Marking technology, the genetic structure and genetic relationship of two clam populations collected in Guangxi with great variation in shape and shell color and Shandong clam population were studied. This document did not provide information on enzyme digestion, pre-amplification, and selective amplification. Reference reaction system

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  • Method for analyzing genetic diversity of shellfish by using fAFLP (Fluorescent Amplified Fragment Length Polymorphism) labeling technology
  • Method for analyzing genetic diversity of shellfish by using fAFLP (Fluorescent Amplified Fragment Length Polymorphism) labeling technology
  • Method for analyzing genetic diversity of shellfish by using fAFLP (Fluorescent Amplified Fragment Length Polymorphism) labeling technology

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Embodiment Construction

[0022] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0023] The advantages of the present invention will be further described below in conjunction with specific examples of silver staining and fluorescent labeling methods for detecting AFLP diversity in different geographical populations of clams.

[0024] (1) Extraction of high-quality DNA: In July 2006, 20 adductor muscles of clams Shandong population (abbreviated as SD) and Guangxi population (abbreviated as GX) were taken, and genomic DNA was extracted with phenol / chloroform method, and 1% agar Sugar gel electrophoresis and ultraviolet spectrophotometer were used to detect the purity and integrity of DNA, and the concentration of DNA in all samples was adjusted to 100ng / μL.

[0025] (2) Restriction endonuclease digestion of DNA: digest with EcoR I and Mse I two endonucleases. The digestion mixture for each DNA sample includes: DNA 100ng, Ms...

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Abstract

The invention relates to a method for analyzing the genetic diversity of shellfish by using a fAFLP (Fluorescent Amplified Fragment Length Polymorphism) labeling technology, which sequentially comprises the following steps of: extracting DNA of shellfish, digesting the DNA by restriction enzyme, carrying out joint connection on the DNA subjected to the enzyme cutting, carrying out PCR ((Polymerase Chain Reaction) pre-amplification and selective amplification, detecting a product and acquiring and analyzing an image and data. Compared with the prior art, the invention has the advantage of being capable of efficiently, stably and accurately analyzing the genetic diversity of the shellfish by adopting a self-creating reaction system combining with the fAFLP labeling technology. In addition, the invention has the advantages of more amplification sites, high polymorphic site proportion and large Nei genetic diversity.

Description

technical field [0001] The invention relates to a method for analyzing the genetic diversity of shellfish, more specifically a method for analyzing the genetic diversity of shellfish by using fAFLP marker technology. Background technique [0002] AFLP (Amplified Fragment Length Polymorphism, amplified fragment length polymorphism) technology is currently widely used and effective genetic diversity detection method, it combines the stability and RAPD (Random Amplified Polymorphism DNA, Random Amplified Polymorphism DNA) technology is highly efficient, and has many advantages such as small amount of DNA required, large amount of marker information, rich polymorphism, high sensitivity, fast and efficient, and reliable data. , and does not need to know the genetic background of the research organism in advance, and is not affected by factors such as tissue and organ types, developmental stages, and habitat conditions. The results follow the characteristics of Mendelian separatio...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 董迎辉林志华
Owner ZHEJIANG WANLI UNIV
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