牛皮革PCR鉴别方法研究  

Identification of Bovine Leather Using PCR Technology

作　　者：柯振华[1,2] 罗海英[1,2] 陈筱婷[1,2] 陈宗良[1,2] 郭新东[1,2] 吴玉銮[1,2] 覃芳芳[1,2] 

机构地区：[1]广州市质量监督检测研究院,广东广州510100 [2]国家皮革制品质量监督检验中心(广州),广东广州510100

出　　处：《皮革科学与工程》2013年第1期32-36,42,共6页Leather Science and Engineering

基　　金：国家质量监督与检验检疫总局科技项目(2009QK372)

摘　　要：本研究比较了不同的DNA提取方法,最终运用苯酚-氯仿抽提法成功地从牛皮革样品中提取获得质量较高的DNA,建立了牛皮革DNA的提取方法。该方法充分考虑了皮革材料的特殊性,利用裂解缓冲液释放皮革中的核酸,采用高浓度盐溶液除去溶液中的蛋白质,用苯酚、氯仿混合溶液进一步除去蛋白,最终获得纯化的皮革DNA。根据牛线粒体基因序列设计特异性的扩增引物,成功地从皮革样品DNA中扩增出271 bp的牛内源基因片段。本方法通过分析皮革样品中的遗传学信息来鉴定皮革种类,特异性强、灵敏度高,可广泛应用于皮革的鉴别与分类中。The total DNA was extracted from leather of bovine and the methods for bovine leather identification based on PCR were established. Considering the special properties of leather materials, in order to DNA of leather the phenol - chloroform extraction method has been selected. In this research, the lysis buffer was used to release the nucleic acid in leather. Then the phenol.- chloroform mixed solution and high - salty solution were applied to remove the protein in the lysis solution. PCR were performed using a pair of universal primers localized in the mitochondrial DNA of cattle. Fragments of about 271bp in size of mitochondrial DNA were amplified by PCR. The genome electrophoresis and PCR experiment proved that we got high - quality I）NA of bovine leather. In general, the PCR method established in this study was a kind of sensitive and specific detection technique for leather identification.

关 键 词：牛皮革 皮革鉴别 脱氧核糖核酸 聚合酶链式反应 

分 类 号：TS57[轻工技术与工程—皮革化学与工程]