酱油、烤鳗酱油DNA提取及原料成分的荧光PCR检测

Extraction of DNA from Soy Sauce and Roast Eel Sauce and Detection of Raw Material Ingredients Using Fluorescence PCR

用4种预处理方法对酱油进行浓缩,后采用CTAB沉淀法提取DNA,再用SYBRGreenⅠ荧光PCR分别检测人工添加的大米gos9基因、酱油原料成分(大豆Lectin基因和小麦Wx012基因)。结果显示:仅人工添加大米DNA、用CTAB沉淀液浓缩的预处理方法提取的酱油DNA中,大米gos9基因、大豆Lectin基因和小麦Wx012基因的检测结果均为阳性,表明这种方法最适于酱油DNA的提取。将建立的DNA提取方法用于3份酱油、4份烤鳗酱油同样有效,gos9基因检测结果均为阳性。2份酱油检出大豆成分,2份烤鳗酱油检出小麦成分,1份酱油和2份烤鳗酱油检出大豆成分和小麦成分。

The soy sauce was concentrated by four pretreatment methods and the subjected to DNA extraction by CTAB precipitation method.SYBR Green Ⅰ fluorescent PCR technique was used to detecting the added artificially gos9 gene in rice,soybean Lectin gene and wheat Wx012 gene in soy sauce raw material ingredients.The results showed that the rice gos9 gene,soybean Lectin gene and wheat Wx012 gene were positive in the DNA extract by the pretreatment method of adding artificially the rice DNA and concentrating by CTAB precipitation solution,which indicated that this method was the most suitable for extracting soy sauce DNA.The DNA extraction method established was employed in analyzing three soy sauce samples and four roast eel sauce samples and results indicated positive of gos9 gene in all samples.The soybean ingredient was detected in two portions of soy sauce,and the wheat ingredient was detected in two portions of roast eel sauce,while the soybean and wheat ingredients were detected in one portion of soy sauce and two portions of roast eel sauce.