Here we present a multiplex PCR assay to distinguish three widespread brown frog species, Rana temporaria, R. arvalis and R. amurensis, based on amplified fragment length. The system consists of four primers to the mitochondrial cytochrome c oxidase subunit I gene, with one universal reverse primer and three species-specific forward primers. Therefore, the PCR gives a fragment of a known specific length, which can be identified by electrophoresis in standard agarose gel. The efficiency of the developed assay was confirmed using both pre-identified and unidentified samples.

中文翻译：

---

多重 PCR 检测可区分三种广泛分布的褐蛙物种

在这里，我们提出了一种多重 PCR 检测方法来区分三种广泛分布的褐蛙物种：林蛙 ( Rana temporaria)、R。阿瓦利斯和R . amurensis，基于扩增片段长度。该系统由线粒体细胞色素 c 氧化酶亚基 I 基因的 4 条引物组成，其中包括 1 条通用反向引物和 3 条物种特异性正向引物。因此，PCR 给出了已知特定长度的片段，可以通过标准琼脂糖凝胶电泳进行鉴定。使用预先鉴定的和未鉴定的样品证实了所开发的测定的效率。