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Molecular marker for identifying carp varieties and method for identifying by utilizing molecular marker

A molecular marker and variety technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as inability to accurately identify carp species

Inactive Publication Date: 2015-09-30
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to overcome the deficiency that morphological characteristics cannot accurately identify carp species, provide a TRAP molecular marker and its obtaining method, which will help to quickly and accurately identify Furui carp and its original parents - Jian carp and Yellow River carp. It has great application value in carp breeding, identification, protection and utilization of germplasm resources

Method used

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  • Molecular marker for identifying carp varieties and method for identifying by utilizing molecular marker
  • Molecular marker for identifying carp varieties and method for identifying by utilizing molecular marker
  • Molecular marker for identifying carp varieties and method for identifying by utilizing molecular marker

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Experimental program
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Effect test

Embodiment 1

[0018] Furui carp and its original parents—Jian carp and Huanghe carp were obtained from the Yixing Experimental Base of the Freshwater Fishery Research Center of the Chinese Academy of Fishery Sciences, and 10 of each fish were randomly selected. Genomic DNA was extracted from the fin rays of each fish.

[0019] GH(GenBank accession no.M27000.1), GHR(AY741100.1), IGF(D83272.1), TRH(AB179818.1) and GTH(X56497.1) genes that may be related to carp growth traits were selected as target candidates Gene, 18 fixed primers were designed. According to the characteristics of exons or introns, four random primers rich in GC or AT core regions were designed and synthesized according to Hu (2006). The details are shown in Table 1.

[0020] Using the genomic DNA as a template, 18 fixed primers were combined with 4 random primers for PCR amplification. The primers are shown in Table 1.

[0021] The total volume of the PCR reaction was 15 μL, where Mg 2+ 1.5mmol / L, dNTPs0.35mmol / L, rando...

Embodiment 2

[0025] Furui carp and one of its original parents, Jian carp, was taken from the Nanquan Experimental Base of Freshwater Fishery Research Center of the Chinese Academy of Fishery Sciences, and the other original parent, the Yellow River carp, was taken from the Yellow River Carp Breeding Farm in Zhengzhou, Henan. Take 10 tails. Genomic DNA was extracted from the fin rays of each fish.

[0026] Using the genomic DNA as a template, 18 fixed primers were combined with 4 random primers for PCR amplification. The primers are shown in Table 1.

[0027] The total volume of the PCR reaction was 15 μL, where Mg 2+ 1.5mmol / L, dNTPs0.35mmol / L, random primer 6pmol / L, fixed primer 10pmol / L, DNA template 60ng, Taq DNA polymerase 1.0U, make up the volume with sterilized double distilled water. The PCR reaction conditions were: 94°C for 4min; 94°C for 45s, 35°C for 45s, 72°C for 1min, 5 cycles; 94°C for 45s, 53°C for 45s, 72°C for 1min, 35 cycles; 72°C for 10min.

[0028] After the PCR rea...

Embodiment 3

[0031] Ten Furui carp and their original parents—7 Jian carp and 7 Huanghe carp were obtained from the Yangtze River Fisheries Research Institute of the Chinese Academy of Fishery Sciences. Genomic DNA was extracted from the fin rays of each fish.

[0032] Using the genomic DNA as a template, 18 fixed primers were combined with 4 random primers for PCR amplification. The primers are shown in Table 1.

[0033] The total volume of the PCR reaction is 15 μl, where Mg 2+ 1.5mmol / L, dNTPs0.35mmol / L, random primer 6pmol / L, fixed primer 10pmol / L, DNA template 60ng, Taq DNA polymerase 1.0U, make up the volume with sterilized double distilled water. The PCR reaction conditions were: 94°C for 4min; 94°C for 45s, 35°C for 45s, 72°C for 1min, 5 cycles; 94°C for 45s, 53°C for 45s, 72°C for 1min, 35 cycles; 72°C for 10min.

[0034] After the PCR reaction, the products were separated by electrophoresis using 8.0% non-denaturing polyacrylamide gel, and after staining by silver staining, pho...

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Abstract

The invention relates to a molecular marker for identifying carp varieties and a method for identifying by utilizing the molecular marker, and belongs to the technical field of molecular markers. A fixed primer sequence and a random primer sequence are provided, and thenPCR reaction is carried out through the pair of primers, so as to obtain the result after electrophoresis. According to the molecular marker for identifying carp varieties and the method for identifying by utilizing the molecular marker, provided by the invention, by adopting the primer pair, a strip of obvious specific band about 200 bp is obtained only in a cyprinus carpio sample through amplification, and is not obtained in the original parent cyprinus carpiovar Jian and Yellow river carp. The molecular marker can be used for identification of carp varieties, breed conservation and breeding assisted by molecular-biological techniques.

Description

technical field [0001] The invention relates to a molecular marker for identifying carp species and a method for identifying them, belonging to the technical field of molecular markers. Background technique [0002] Carp is one of the most important freshwater aquaculture species in my country, and there are many species of aquaculture. At present, there are more than 20 carp species that have obtained the certificate of new aquatic species. When distinguishing different carp species, it is often based on morphological characteristics, such as body color or body length, body height, lateral line scales, fin rays and other countable and measurable traits. Different carp species have partial overlap, so that it is impossible to accurately distinguish different species. [0003] Molecular markers are genetic markers based on nucleotide sequence variation in genetic material between individuals, and are a direct reflection of genetic polymorphism at the DNA level. Among them, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 董在杰曲疆奇朱文彬王兰梅
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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