Molecular genetic methods are increasingly used to supplement or substitute classical morphology-based species identification. Here, we employ a COI mini-barcode coupled high-resolution melting analysis to quickly, cost-efficiently and reliably determine larvae of two closely related Cychramus (Coleoptera, Nitidulidae) species. Euclidean distance comparison (p < 0.01) and a Welch t-test of the melting point temperatures (p < 0.01) provide highly significant statistical evidence for species specific differences in melting and fluorescence curves, thus allowing the assignment of larvae to either of the two species. This protocol serves as a fast, low-cost and low-tech method to discriminate between pairs or groups of closely related species and can be adapted and applied to various ecological research questions.

分子遗传学方法越来越多地用于补充或替代基于经典形态学的物种识别。在这里，我们采用COI微型条形码耦合的高分辨率熔解分析来快速，经济高效且可靠地确定两个密切相关的Cycyramus（鞘翅目，Nitidulidae）物种的幼虫。欧几里德距离比较（p <0.01）和熔点的Welch t检验（p <0.01）为物种特定的熔解和荧光曲线差异提供了非常重要的统计证据，因此可以将幼虫分配给这两个中的任何一个种类。该协议是一种快速，低成本和低技术的方法，可用于区分成对的或紧密相关的物种，并且可以适用于各种生态研究问题。