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Duplex PCR method for identifying genetic relationship of black carp

A kind of kinship and herring technology, applied in the field of animal molecular genetics, has achieved great practical value, reduced time and cost, and high polymorphism

Active Publication Date: 2019-03-15
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no report on multiplex PCR of microsatellite markers in herring at home and abroad

Method used

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  • Duplex PCR method for identifying genetic relationship of black carp
  • Duplex PCR method for identifying genetic relationship of black carp
  • Duplex PCR method for identifying genetic relationship of black carp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Design 1,000 pairs of microsatellite marker primers through transcriptome sequencing, use herring DNA as a template, test the quality of these 1,000 pairs of microsatellite marker primers, screen clear bands and high polymorphism microsatellite marker primers, and finally obtain 52 pairs of markers Microsatellite markers with high morphological properties and stable amplification. Then, according to the size of the PCR product amplified by the microsatellite marker primers obtained by screening, pairwise pairing was performed to screen for combinations with high stability, and finally a double PCR reaction system of 5 groups of herring microsatellite markers was obtained. The primers are as follows:

[0020]

Embodiment 2

[0022] A practical application of a double PCR system in the identification of herring phylogenetic relationship, the steps are:

[0023] 1. Randomly select 30 herrings from the 4 herring populations in the herring farm. Take one fish from the first population, 2 fish from the second population, 3 fish from the third population, and 24 fish from the fourth population. Each fish was marked and marked, and then the fin rays of the herring were collected. In order to prevent human intervention in the experimental results, the order of the samples was disrupted, and the accuracy of the method was verified according to the experimental results. The order after scrambling is: the 9th fish is taken from the first population, the 1st and 11th fish are from the second population, the 10th, 13th and 19th fish are from the third population, and the rest are from the fourth population populations. Genomic DNA of herring is extracted, and the extraction method adopts the kit extraction me...

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Abstract

The invention provides a duplex PCR method for identifying the genetic relationship of black carp. The method comprises the steps as follows: (1) extracting genome DNA of a black carp sample; (2) performing duplex PCR by 5 groups of microsatellite primers; (3) performing electrophoretic separation on a PCR amplified product on polyacrylamide gel; (4) counting genotype of the PCR amplified productand analyzing the genetic relationship between individuals. The 5 groups of microsatellite primers are sites with high polymorphism and clear strips and can realize genetic diversity detection and individual recognition of the black carp and identify the genetic relationship between individuals, inbreeding among black carp individuals can be avoided, and a the used time and cost are a half of those in traditional PCRs. Tests prove that the method can identify the genetic relationship of 30 black carps and accurately identify the genetic relationship between samples, and the genetic distances between the samples can be calculated according to identification results.

Description

technical field [0001] The invention belongs to the field of animal molecular genetics, and in particular relates to a double PCR method applied to identification of herring kinship. Background technique [0002] Herring (Latin scientific name: Mylopharyngodon piceus; English name: Black carp), Osteostichthyes, Radiopteridae, Cypriniformes, Cyprinidae, Arrow subfamily, Herring genus. Herring, also known as black carp, black carp, and snail carp, is commonly known as "black carp", "black carp" or "snail carp" in Anhui. It is named for its black body and preference for eating snails. [0003] Body color black, fin gray black. The head is wide and flat, the mouth is terminal, without beards, and the lower pharyngeal teeth are 1 row, which are molar-shaped. Inhabits the middle and lower layers, and feeds mainly on snails, mussels, shrimps and aquatic insects. Distributed in major water systems in China, mainly in the plains south of the Yangtze River in my country, and rare i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/16
Inventor 刘明典宋江腾汪登强段辛斌刘绍平陈大庆
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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