应用多重PCR同时检测牛、羊源性成分

Applies Multiplex PCR Simultaneously to Examine the Bovine and Sheep Derived Materials

我们选择了3对引物A,B,C。A可扩增大多数脊椎动物(哺乳类、鸟类、爬行类、两栖类和鱼类)细胞色素b基因上一个371bp的区段,B可扩增位于牛mtDNAD-loop区段内的一个274bp的区段,C可扩增羊mtDNA基因上一个199bp的区段。由于3对引物扩增的产物分别相差约100bp左右,可通过琼脂糖凝胶电泳分离来并观察结果,建立了基于内对照的同时检测牛、羊源性成分的三重PCR方法。

Three pairs of primers, A, B and C, were selected for multiplex PCR detection. A fragment of 371bp on cytochrome b gene of the most vertebrate animals could be amplified from primers A, a fragment of 274bp on the loop region of bovine mitochondria DNA (mtDNA) from primers B and a fragment of 199bp on the sheep mitochondria gene from primers C. As the three amplification products were different in length, the result could be easily observed by agarose gel electrophoresis. In conclusion, triple PCR on the basis of internal control was established for simultaneous detection of bovine and Sheep derived materials.