基于coⅰ基因鹌鹑肉源成分pcr扩增鉴定方法和专用引物

COI (oxidase subunit I) gene based quail-meat-derived ingredient PCR (polymerase chain reaction) amplification method and special primer

Abstract

The invention discloses a method for quickly detecting quail-derived ingredients in food and a special primer. The special primer is shown in a sequence table SEQ ID NO 1-2. According to the method, PCR (polymerase chain reaction) and qPCR (quantitative polymerase chain reaction) are performed on a mixed template in which a quail-meat-derived DNA serves as a positive template and DNAs of cattle, sheep, pigs, rabbits, pigeons, chicken, ducks and gooses serve as interference templates. An electrophoretic band in a agarose gel electrophoretogram serves as a positive result of a PCR method, amplification of a Ct value, smaller than 34, of an amplification curve serves as a positive result of a qPCR method, and detection sensitivity reaches a peak-level DNA. The method has the advantages that a COI gene is used as a target gene to detect deep processed quail meat for the first time, the designed primer is applicable to the PCR method and the qPCR method, high specificity is achieved, a new approach to identification of animal-derived ingredients in the food is created due to high sensitivity, the method is convenient to operate, stable and accurate, and the like.
本发明公开了一种快速检测食品中鹌鹑源性成分的方法及专用引物。专用引物见序列表SEQ ID NO 1‑2。针对以鹌鹑肉源DNA为阳性模板,牛、羊、猪、兔、鸽、鸡、鸭和鹅8个物种的DNA为干扰模板的混合模板,进行PCR和qPCR反应。其中PCR方法的结果以琼脂糖凝胶电泳图谱中电泳条带为阳性结果,而qPCR方法中以扩增曲线的Ct值小于34的扩增作为阳性结果,检测灵敏度达皮克级DNA。该方法首次利用COⅠ基因为目标基因对深加工鹌鹑肉进行检测,所设计引物同时适用于PCR和qPCR方法中,不仅具有良好的特异性,且高度的灵敏性为食品中动物源性成分的鉴定探索了新的途径,具有操作便捷,稳定准确等有益结果。

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