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目的:建立肝水解肽各工艺步骤段中样品的PCR检测方法,用于检测生产原料的动物来源是牛源性或猪源性。方法:分别对猪源性及牛源性肝水解肽各工艺步骤段中样品进行了DNA提取,并加入猪、牛特异性引物进行扩增。对扩增产物进行酶切及测序验证。结果:工艺段为超滤液四之前的样品均可提取并扩增出相应的DNA片段,牛源性PCR产物与Genbank中牛序列的同源性为100%,猪源性PCR产物与猪序列的同源性为99.4%,PCR扩增的检测限为0.5 mg·mL-1的肝细胞酶解液。结论:可用本方法检测肝水解肽各工艺步骤段中超滤液四之前的样品。
OBJECTIVE: To establish a PCR method for the detection of samples in each step of the process of hepatic hydrolyzate. The animal origin for the production of raw materials is bovine or porcine origin. Methods: DNA samples were extracted from each step of the process of pig-originated and bovine liver-derived peptide. The pigs and cattle-specific primers were used for amplification. The amplified product was digested and sequenced. Results: The corresponding DNA fragment was extracted and amplified from the sample before the fourth ultrafiltrate. The homology between the bovine source PCR product and the cow sequence in Genbank was 100% The homology was 99.4%. The detection limit of PCR amplification was 0.5 mg · mL-1. Conclusion: This method can be used to detect hepatic hydrolytic peptide in each step of the ultrafiltrate step four samples.