| Identifying tissue DNA of Far Eastern sardine or Japanese sardine in e.g. fish product, by performing DNA extraction from animal tissue, setting up PCR with fluorescent detection, measuring accumulation of fluorescent signals, and interpreting results | |
| 2023-06-23 | |
| 专利权人 | UNIV KUBAN STATE AGRARIAN (UYKU-Non-standard) |
| 申请日期 | 2023-06-23 |
| 专利号 | RU2023116697-A; RU2835037-C2 |
| 成果简介 | NOVELTY - Method for identifying tissue DNA of Far Eastern sardine or Japanese sardine (Sardinops melanostictus) in fish product, or in fishmeal and bone meal in feed involves performing DNA extraction from animal tissue by sorption method, setting up a PCR with fluorescent detection with 45 real-time amplification cycles using oligonucleotide primers, probes and fluorescent dyes specific to region of animal DNA genome, measuring accumulation of fluorescent signals through channels of corresponding fluorescent dyes, and interpreting results based on the presence or absence of intersection of the fluorescence curve with the threshold line, where if the fluorescence signal accumulation curves appear before the 35th cycle, the reaction result is considered positive, and if the curves do not cross the threshold line or cross it after the 35th cycle, the reaction result is negative. USE - The method is useful for identifying tissue DNA of Far Eastern sardine or Japanese sardine in fish product, or in fishmeal and bone meal in feed. DETAILED DESCRIPTION - Method for identifying tissue DNA of Far Eastern sardine or Japanese sardine (Sardinops melanostictus) in fish product, or in fishmeal and bone meal in feed involves performing DNA extraction from animal tissue by sorption method, setting up a PCR with fluorescent detection with 45 real-time amplification cycles using oligonucleotide primers, probes and fluorescent dyes specific to region of animal DNA genome, measuring accumulation of fluorescent signals through channels of corresponding fluorescent dyes, and interpreting results based on the presence or absence of intersection of the fluorescence curve with the threshold line, where if the fluorescence signal accumulation curves appear before the 35th cycle, the reaction result is considered positive, and if the curves do not cross the threshold line or cross it after the 35th cycle, the reaction result is negative. In method, JOE(RTM: 2',7'-Dimethoxy-4',5'-dichloro-6-carboxyfluorescein)/Yellow and CY5(RTM: Cyanine 5)/Red is used for animal-specific signal, a suspension of bacteriophage T4 with a concentration of 5x 103phage particles/1 μ l is used as an internal control sample, and a mixture containing fragments of animal genome and bacteriophage T4 with nucleotide sequence: T4F tacatataaatcacgcaaagc (SEQ ID NO: Not defined) (forward primer), T4R tagtatggctaatcttattgg (SEQ ID NO: Not defined) (reverse primer) and T4P CY5 acattggcactgaccgagttc (SEQ ID NO: Not defined) (probe) in a ratio of 1:1 is used as a positive control sample. The DNA is isolated from tissue of the Far Eastern sardine or Japanese sardine, and a fragment of the genome of the tissue of the Far Eastern sardine or Japanese sardine with nucleotide sequence is used for positive control sample, preferably Sm-F forward primer having a base pair sequence: ggtagagcaattccctgcgt (SEQ ID NO: Not defined), Sm-R reverse primer having a base pair sequence: ccgagtcccctcatggtttt (SEQ ID NO: Not defined) and Sm-Z probe having a base pair sequence: R6G-cattggactattaggacccgca-Black Hole Quencher-1(RTM: Not defined) (SEQ ID NO: Not defined). |
| 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/21527 |
| 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 tissue DNA of Far Eastern sardine or Japanese sardine in e.g. fish product, by performing DNA extraction from animal tissue, setting up PCR with fluorescent detection, measuring accumulation of fluorescent signals, and interpreting results. RU2023116697-A; RU2835037-C2[P]. 2023. |
| 条目包含的文件 | 条目无相关文件。 | |||||
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