Data from: Sex determination in the wild: a field application of loop-mediated isothermal amplification successfully determines sex across three raptor species

PCR-based methods are the most common technique for sex determination of birds. Although these methods are fast, easy and accurate, they still require special facilities that preclude their application outdoors. Consequently, there is a time lag between sampling and obtaining results that impedes researchers to take decisions in situ and in real time considering individuals’ sex. We present an outdoor technique for sex determination of birds based on the amplification of the duplicated sex-chromosome-specific gene Chromo-Helicase-DNA binding protein using a loop-mediated isothermal amplification (LAMP). We tested our method on Griffon Vulture (Gyps fulvus), Egyptian Vulture (Neophron percnopterus) and Black Kite (Milvus migrans) (family Accipitridae). We introduce the first fieldwork procedure for sex determination of animals in the wild, successfully applied to raptor species of three different subfamilies using the same specific LAMP primers. This molecular technique can be deployed directly in sampling areas because it only needs a voltage inverter to adapt a thermo-block to a car lighter and results can be obtained by the unaided eye based on colour change within the reaction tubes. Primers and reagents are prepared in advance to facilitate their storage at room temperature. We provide detailed guidelines how to implement this procedure, which is simpler (no electrophoresis required), cheaper and faster (results in c. 90 min) than PCR-based laboratory methods. Our successful cross-species application across three different raptor subfamilies posits our set of markers as a promising tool for molecular sexing of other raptor families and our field protocol extensible to all bird species.

基于聚合酶链式反应（PCR）的方法是当前鸟类性别鉴定最常用的技术手段。尽管此类方法具备快速、简便且准确的优势，但仍需专用实验设备，这使其无法在野外环境中开展应用。因此，采样与获取结果之间存在时间延迟，这阻碍了研究人员在野外现场结合个体性别状况实时做出决策。
本研究提出一种基于环介导等温扩增（LAMP）技术、通过扩增性别染色体特异性重复基因Chromo-螺旋酶-DNA结合蛋白（Chromo-Helicase-DNA binding protein）的鸟类性别鉴定户外方案。本研究对秃鹫（Gyps fulvus）、白兀鹫（Neophron percnopterus）以及黑鸢（Milvus migrans，隶属鹰科Accipitridae）开展了方法验证。本研究首创了适用于野生动物野外性别鉴定的作业流程，并利用同一套特异性LAMP引物，成功将其应用于三个不同猛禽亚科的物种。
该分子技术可直接在采样区域部署：仅需一台电压转换器，即可将恒温金属浴适配至车载点烟器供电；反应管内的颜色变化可通过肉眼直接读取结果。引物与反应试剂可提前配制完成，便于在室温条件下储存。本研究提供了该流程的详细实施指南：相较于基于PCR的实验室方法，该方案无需电泳步骤，操作更简便、成本更低廉且耗时更短（约90分钟即可获得结果）。
本研究成功在三个不同猛禽亚科间实现跨物种应用，表明这套标记可作为其他猛禽科物种分子性别鉴定的潜在工具，且本野外方案可推广至所有鸟类物种。