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
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[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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