Identifying Cololabis saira tissue DNA in fish products, in fishmeal and feed, by DNA extraction from animal tissue using the sorption method, polymerase chain reaction with fluorescent detection with real-time amplification cycles
	CHERNYKH O YU; SHEVCHENKO A A; KOSHCHAEV A G; KRIVONOS R A; SHEVCHENKO L V; MANAKOVA A I; YAKOVLENKO P P; AGOLTSOV V A; SHEVCHENKO A N; BANNOV V A; CHERNOV A N; MALYSHEV D V; ZABASHTA S N; BELOUSOV V I; ADIATULIN I F; SEMENENKO M P
	2023-06-23
专利权人	UNIV KUBAN STATE AGRARIAN (UYKU-Non-standard)
申请日期	2023-06-23
专利号	RU2023116698-A
成果简介	NOVELTY - Identifying Pacific saury (Cololabis saira) tissue DNA in fish products, in fishmeal and feed, includes DNA extraction from animal tissue using the sorption method, polymerase chain reaction with fluorescent detection with 45 real-time amplification cycles using oligonucleotide primers, probes, and fluorescent dyes specific to the animal's DNA genome region for the animal-specific signal-JOE/Yellow and Cy5/Red for the internal control sample in the form of a suspension of bacteriophage T4 with a concentration of 5x 103phage particles per 1 μ l, a positive control sample in the form of a mixture containing fragments of animal genomes and bacteriophage T4 with the nucleotide sequence T4F TACATATAAATCACGCAAAGC- forward primer, T4R TAGTATGGCTAATCTTATTGG-reverse primer, T4P CY5 ACATTGGCACTGACCGAGTTC- probe, taken in a 1:1 ratio and measuring the accumulation of fluorescent signals in the channels of the corresponding fluorescent dyes. USE - The method is useful for identifying Pacific saury (Cololabis saira) tissue DNA in fish products, in fishmeal and feed. ADVANTAGE - None given. DETAILED DESCRIPTION - Identifying Pacific saury (Cololabis saira) tissue DNA in fish products, in fishmeal and feed, includes DNA extraction from animal tissue using the sorption method, polymerase chain reaction with fluorescent detection with 45 real-time amplification cycles using oligonucleotide primers, probes, and fluorescent dyes specific to the animal's DNA genome region for the animal-specific signal-JOE/Yellow and Cy5/Red for the internal control sample in the form of a suspension of bacteriophage T4 with a concentration of 5x 103phage particles per 1 μ l, a positive control sample in the form of a mixture containing fragments of animal genomes and bacteriophage T4 with the nucleotide sequence T4F TACATATAAATCACGCAAAGC- forward primer, T4R TAGTATGGCTAATCTTATTGG-reverse primer, T4P CY5 ACATTGGCACTGACCGAGTTC- probe, taken in a 1:1 ratio and measuring the accumulation of fluorescent signals in the channels of the corresponding fluorescent dyes, interpreting the results based on the presence or absence of intersection of the fluorescence curve with the threshold line, if the fluorescent signal accumulation curves appear before the 35th cycle, then the reaction result is considered positive, and if the curves do not cross the threshold line or cross it after the 35th cycle, then the result of the reaction is negative, differing in that DNA is isolated from the tissue of the Pacific saury and for the positive control sample a fragment of the genome of the tissue of the Pacific saury with the nucleotide sequence having Cs-F AGGGACAAAAATCGTGGGGG primer, Cs-RCGTGTGCACTCTGAAATGCC primer and Cs-ZR6GCATGGTGGGTGACAGGGTAA BHQ1 probe.
IPC 分类号	C12Q-001/68
国家	俄罗斯
专业领域	生物科学
语种	英语
成果类型	专利
文献类型	科技成果
条目标识符	http://119.78.100.226:8889/handle/3KE4DYBR/21526
专题	中国科学院新疆生态与地理研究所
作者单位	UNIV KUBAN STATE AGRARIAN (UYKU-Non-standard)
推荐引用方式 GB/T 7714	CHERNYKH O YU,SHEVCHENKO A A,KOSHCHAEV A G,et al. Identifying Cololabis saira tissue DNA in fish products, in fishmeal and feed, by DNA extraction from animal tissue using the sorption method, polymerase chain reaction with fluorescent detection with real-time amplification cycles. RU2023116698-A[P]. 2023.

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