CH4 enzyme digestion identifying method of hybrid yellow catfish

An enzyme digestion identification and yellow catfish technology, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of low detection efficiency, low repeatability, and influence on popularization and use, and achieves simple operation, experimental Fast, intuitive identification of results

Active Publication Date: 2017-01-04
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Afterwards, Zhao Wenxue et al. (RAPD molecular identification and genetic analysis of hybrids of Peeled catfish species [J]. 2006.30 (1): 101-105) carried out the germplasm identification of Peeled catfish and its hybrids. However, the identification The RAPD marker used in the work is limited by the method itself, and the detection efficiency is low and the repeatability is not high, which seriously affects the promotion and use of this method.

Method used

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  • CH4 enzyme digestion identifying method of hybrid yellow catfish
  • CH4 enzyme digestion identifying method of hybrid yellow catfish
  • CH4 enzyme digestion identifying method of hybrid yellow catfish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] CH 4 Enzyme digestion identification method, using CH after PCR amplification 4 There are interspecies sequence differences between the gene fragments of Pedelvis catfish and P. Pelvic catfish and its hybrid offspring.

Embodiment 2

[0030] CH 4 Enzyme digestion identification method specifically comprises the following steps:

[0031] A, take the tissue sample of the fish to be tested, and extract the genomic DNA;

[0032] B. Using the extracted genomic DNA as a template, carry out PCR amplification under the guidance of special primers;

[0033] C. Digest the amplified target gene fragment with restriction endonuclease HindⅢ, and the restriction site is A↓AGCTT;

[0034] D. PCR-RFLP product was detected by agarose gel electrophoresis. After electrophoresis detection, there was only one band on the same swimming lane, which was Peeled catfish; there were two bands on the same swimming lane, which was Peeled catfish; in the same swimming lane There are three bands on it, which is a hybrid.

Embodiment 3

[0036] CH 4 Enzyme digestion identification method specifically comprises the following steps:

[0037] A, take the tissue sample of the fish to be tested, and extract the genomic DNA;

[0038] B. Using the extracted genomic DNA as a template, carry out PCR amplification under the guidance of special primers;

[0039] C. Digest the amplified target gene fragment with restriction endonuclease HindⅢ, and the restriction site is A↓AGCTT;

[0040] D. The PCR-RFLP product was detected by agarose gel electrophoresis. After electrophoresis detection, there was only one band on the same swimming lane, which was peltatus varschii.

[0041] The special primer in the B step is CH 4 Specific primers for gene fragments, the primer pair is:

[0042] Forward primer 5'-TCCCCAAGGTTTACTTGCTCGCTCC-3';

[0043] Reverse primer 5'-CGATGGATCTGGATATGTGGCGCAC-3'.

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Abstract

The invention provides a CH4 enzyme digestion identifying method of hybrid yellow catfish. The CH4 gene segment after PCR amplification has the interspecific crossing sequence difference between the yellow catfish and pelteobagrus vachelli; the restriction enzyme Hind III is applied to enzyme digestion; the enzyme digestion locus is A->AGCTT; the yellow catfish, pelteobagrus vachelli and hybrid descendants of the yellow catfish and pelteobagrus vachelli are analyzed and identified by application of a RFLP (restriction fragment length polymorphism) method. The hybrid and the purebred which are not easy to distinguish morphologically can be distinguished from the level of molecular biology; the method is fast in experiment, simple in operation, direct in identification and free from sequence testing.

Description

technical field [0001] The present invention relates to the identification method of fish hybrid, in particular to the CH of aquaculture yellow catfish hybrid 4 Molecular marker identification method. Background technique [0002] The fish of the genus Pelida belongs to omnivorous demersal fish, mainly carnivorous, and is one of the most developed teleosts in fish evolution. Both Pelteobagrus fulvidraco and Pelteobagrus vachelli belong to the Siluriformes, Paleidae, and Pelteobagrus genus. Both are delicious and have high nutritional value, and are important farmed fish. Yellow catfish are distributed in the Yangtze River, Yellow River, Pearl River, Heilongjiang, Liaoning and other river basins in my country. They like to inhabit open water bodies and live at the bottom. Wantuo and river bottom gravel river sections are suitable for them to inhabit. The 1-2 year old fish grow faster. Fast, slow growth in the future, its individual is small, but its ability to adapt to the e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2521/301C12Q2531/113
Inventor 文秦超荆慧芳黎树张修月宋昭彬
Owner SICHUAN UNIV
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