羊皮革PCR鉴别方法研究

Identification of Sheep Leather Using PCR Technology

针对羊革,运用苯酚-氯仿2次纯化法成功地从羊革样品中获得质量相对较高的DNA,建立了羊革PCR的鉴定方法。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,采用苯酚、氯仿混合溶液2次纯化的方法除去蛋白,最终获得纯化的皮革DNA。根据羊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294bp的羊内源基因片段,检测率达到62.5%。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、可作为皮革材质鉴别的一种手段。

The high quality of sheep leather was successfully purified by the methods of phenol - chloroform secondary purifica- tion with PCR method. Fully taken account the particularity of leather material, the lysis buffer was used to release the nucleic acid in leather, then the phenol -chloroform mixed solution were twice applied to remove the protein in the lysis solution. PCR were performed using a pair of universal primers localized in the mitochondrial DNA of sheep. Fragments of about 294bp in size of mitochondrial DNA are amplified by PCR, the detection rate is 62.5 %. In general, the PCR method established is a kind of spe- cific detection technique for sheep leather identification.