中文摘要：本发明公开了一种转基因鲑鱼AquAdvantage品系特异性实时荧光PCR检测引物、检测方法和试剂盒。本发明针对转基因鲑鱼AquAdvantage品系特异性序列(美洲绵鳚抗冻蛋白基因启动子区域与鲑鱼基因组5’端邻接区序列)设计引物和TaqMan探针，建立转基因鲑鱼实时荧光聚合酶链式反应(polymerase?chain?reaction，PCR)检测方法，利用该检测引物和检测探针，按照本发明的方法进行检测发现，其具有特异性强，灵敏度高，检测灵敏度为60拷贝，检测重复性良好，可应用于转基因鲑鱼AquAdvantage的鉴定，满足监管部门对其转基因鲑鱼正确标识的需求。
外文摘要：NOVELTY - A primer for specific real-time fluorescent PCR detection of transgenic salmon AquAdvantage strain, is new.
USE - The primer is useful in kit and method for specific real-time fluorescent PCR detection of transgenic salmon AquAdvantage strain (all claimed).
ADVANTAGE - The primer detects transgenic salmon AquAdvantage strain with high specificity, and sensitivity, and good repeatability.
DETAILED DESCRIPTION - A primer for specific real-time fluorescence PCR detection of transgenic salmon AquAdvantage strain, is new. The primer comprises Aqu-F comprising base pair sequence of: 5'-cacgttgtactgctgata-3' (SEQ ID NO: Not defined), and Aqu-R comprising base pair sequence of: 5'-gtggtttagttctcatacatc-3' (SEQ ID NO: Not defined). INDEPENDENT CLAIMS are included for the following:
(1) probe comprising base pair sequence of: 5'-cctcacgttgtactactgataccaca-3', where 5' end of the probe is labeled with fluorescent reporter group and 3' end of the probe is labeled with fluorescent quenching group;
(2) kit comprising real-time fluorescent quantitative PCR reaction solution, heat-resistant DNA polymerase, detection primer and detection probe; and
(3) method for real-time fluorescent PCR detection of transgenic salmon AquAdvantage strain, involves (a) extracting DNA sample as template, and adding detection primer and detection probe, heat-resistant DNA polymerase, and real-time fluorescent quantitative PCR reaction solution, and performing real-time fluorescent quantitative PCR and determining whether the sample is transgenic salmon AquAdvantage strain based on the amplification curve, or (b) extracting DNA sample as template, and adding detection primer and detection probe, heat-resistant DNA polymerase, and real-time fluorescent quantitative PCR reaction solution, reacting mixture, obtaining initial concentration of the template corresponding to the cycle threshold (Ct) value, linearly relating Ct value with logarithm value for the initial concentration, and obtaining standard curve for transgenic salmon AquAdvantage strain, performing real-time fluorescent quantitative PCR, and determining whether the sample is transgenic salmon AquAdvantage strain based on the amplification curve.
TF   TECHNOLOGY FOCUS - BIOTECHNOLOGY - Preferred Biomolecule: The fluorescent reporter group is 6-carboxyfluorescein (FAM) and the fluorescent quenching group is black hole quencher 1 (RTM: Not defined).
主权项：一种转基因鲑鱼AquAdvantage品系特异性实时荧光PCR检测引物，其特征在于，所述的检测引物如下所示：Aqu?F：5‘?CACGTTGTACTGCTGATA?3’；Aqu?R：5‘?GTGGTTTAGTTCTCATACATC?3’。
申请号：CN201610855072.3
专利号：CN106282377-A
公开/公告号：CN106282377-A
申请/专利权人：黄埔出入境检验检疫局综合技术服务中心
发明/设计人：刘二龙;吕英姿;卢丽;许龙岩;蒋湘;李立霞;李嘉琪;杜雅萍;林学勤;郑高彬;樊武疆;林先准;李培深;苏彩珠;唐涌
分类号：C12Q1/68;C12N15/11
主分类号：C12Q1/68
法律状态：实质审查
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