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Method for classifying ctenopharyngodon idella based on expressed sequence tag-simple sequence repeats (EST-SSR) marker

A classification method and grass carp technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of insufficient research on the genetic structure of grass carp populations.

Inactive Publication Date: 2011-11-16
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

At present, scholars have used isoenzyme, mtDNA-RFLP, microsatellite and other detection techniques to study the genetic structure of grass carp populations in the Yangtze River system, but there are not many studies on the genetic structure of grass carp populations in different water systems

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  • Method for classifying ctenopharyngodon idella based on expressed sequence tag-simple sequence repeats (EST-SSR) marker
  • Method for classifying ctenopharyngodon idella based on expressed sequence tag-simple sequence repeats (EST-SSR) marker
  • Method for classifying ctenopharyngodon idella based on expressed sequence tag-simple sequence repeats (EST-SSR) marker

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Embodiment Construction

[0025] The applicant constructed a cDNA library of grass carp brain, muscle, liver and other tissues in 2009, obtained 45,318 EST sequences after de-redundancy processing, with an average sequence length of 650bp, and found them using trf (tandem repeats finder) software 5 556 microsatellites. Select a double-base repeat sequence with a repeat number of more than 5 times and a three-base or four-base repeat sequence with a repeat number of more than 4 times, use the primer design software Primer Primer5.0 to design primers, and obtain 118 pairs of EST-SSR primers .

[0026] The applicant used 118 pairs of EST-SSR primers to perform PCR amplification on 50 grass carp samples, of which 87 pairs of primers amplified a single band, 12 pairs of primers had no amplification products, and 19 pairs of primers could amplify polymorphisms Fragments, polymorphic primers accounted for 16.00% of all designed primers. Among them, there are 15 double-base repeat sequences, accounting for 8...

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Abstract

The invention discloses a method for classifying ctenopharyngodon idella based on an expressed sequence tag-simple sequence repeats (EST-SSR) marker, which comprises the following steps of: extracting the deoxyribonucleic acid (DNA) of the ctenopharyngodon idella, amplifying by using the EST-SSR marker, and analyzing the genetic structure of the ctenopharyngodon idella according to an amplification result. Compared with the conventional method, the method has the advantages of high purposiveness and direct action effect, and is easy to operate, high in detection speed, low in detection cost, and convenient to widely popularize and use.

Description

technical field [0001] The invention relates to a fish classification method, in particular to a grass carp classification method based on EST-SSR markers. Background technique [0002] grass carp (Ctenopharyngodon idella) It has the advantages of fast growth and low breeding cost. It is the main breeding object in ponds, lakes and reservoirs, and is one of the most important breeding species in my country's freshwater fishery. Grass carp is distributed in all major river systems in Asia, from Heilongjiang to Xijiang in China (except for Xinjiang and the Qinghai-Tibet Plateau where there is no natural distribution), forming a complex genetic background and rich germplasm resources of grass carp. In recent years, the changes and pollution of the natural water environment have caused a significant decline in the number of natural grass carp populations, and inbreeding in production has led to germplasm degradation, which has affected the normal development of the grass carp a...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 于凌云白俊杰王解香樊佳佳全迎春李胜杰马冬梅叶星
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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