Roe and fry quick classification and determination methods based on CoI DNA bar codes

A technology of rapid classification and identification method, applied in the field of classification and identification of fish eggs and fry, can solve the problems of high cost of classification and identification, difficult amplification of feature fragments, limited degree of discrimination, etc., and achieve short experimental period, low sample requirements, and overcome limitations sexual effect

Inactive Publication Date: 2016-02-24
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

The author found in practice and consulting a large number of literatures that the 16SrDNA sequence corresponding to 16SrRNA is usually used as a molecular marker for microbial community structure and phylogenetic evolution, and 16sRNA has limited discrimination for fish, especi

Method used

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Embodiment 1

[0020] The present invention is a kind of rapid classification and identification method of fish roe and fry based on CoIDNA barcode, adopting the following steps:

[0021] 1. Obtain fish samples that can be accurately classified and identified through fixed-point fishing and market purchases in the surveyed waters, and use 95% analytical pure alcohol to fix them and bring them back to the laboratory. The above fish samples were classified and identified one by one. Take an appropriate amount of dorsal muscle (or fin rays) to extract DNA by conventional phenol / chloroform method, and store it at -20°C for future use.

[0022] 2. Use the DNA obtained in step 1 as a template to amplify the mitochondrial CoI fragment. Amplification primers are universal primers: LC01490: 5'-GGTCAACAAATCATAAAGATATTGG-3', HC02198: 5'-TAAACTTCAGGGTGACCAAAAAATCA-3'.

[0023] 3. The reaction conditions for PCR amplification are: pre-denaturation at 94°C for 3 minutes; each cycle includes 94°C for 40s...

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Abstract

The invention discloses roe and fry quick classification and determination methods based on CoI DNA bar codes. The methods comprise the following steps: collecting fish samples capable of being subjected to accurate classification and determination in a water region under investigation, respectively carrying out PCR amplification and sequencing on mitochondria CoI fragments of various fishes, establishing a CoI DNA bar code database of the fishes in the water region under investigation according to the amplification and sequencing results, numbering the roe and fry samples in the water region under investigation, respectively extracting DNAs of the roe and fry samples, carrying out amplification and sequencing on feature fragments, comparing the amplification and sequencing results with the established CoI DNA bar code database, and thus rapidly and accurately determining the varieties of the roe and fry samples. With the adoption of the methods, the defects that the existing classification and determination methods are large in error and fail in identification, samples to be classified need to be cultured in large quantities, genetically close species can not be distinguished, and the classification and determination cost are high are overcome, and the advantages of rapidness and accuracy of the roe and fry quick classification and determination methods are exerted.

Description

technical field [0001] The invention belongs to the field of fish molecular biology, and in particular relates to a method for classifying and identifying fish eggs and fry. Background technique [0002] Fish eggs and fry are important stages in the early life history of fish. Carrying out early fish resources surveys including fish eggs and fry can not only estimate the number of fish breeding populations, but also estimate the number of supplementary populations, and then scientifically predict changes in fish populations, providing a basis for the protection and rational development and utilization of fishery resources. scientific basis. [0003] The traditional method of identifying fish eggs and fry mainly uses an optical microscope (dissecting mirror) to observe their morphological characteristics. However, in the life history of fish, the morphological changes of fish eggs and fry are complicated, and there are few morphological features that can be used for identif...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888
Inventor 张敏莹徐东坡方弟安周彦锋刘凯段金荣
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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