Environmental DNA (eDNA) is an emerging approach for detecting species, yet numerous methodological questions remain unanswered. Here we examined how time to filtration (0–48 h after collection) and sample storage (open vs. chilled in the dark) influenced detection and measured eDNA concentration. Water samples kept in the dark and chilled had no significant decrease in detection or eDNA concentration relative to those filtered immediately upon return from the field. Water samples exposed to light and ambient air temperature had non-detections beginning at 1 h, the majority of these samples were below the limit of detection (LOD) by 6 h, and eDNA was undetectable in these samples by 24 h. These results have important implications for eDNA research where immediate access to refrigeration is not available, or for fieldwork that requires extended sampling time (e.g., canoeing a river). Further, we report faster eDNA degradation times under ambient conditions than some previous aquaria or mesocosm studies, suggesting an ongoing need to study mechanisms related to eDNA persistence and sample storage.

环境DNA（eDNA）是一种检测物种的新兴方法，但是许多方法学问题仍未得到解答。在这里，我们研究了过滤时间（收集后0-48小时）和样品存储时间（开放与黑暗中冷藏）如何影响检测和测得的eDNA浓度。相对于从田间返回后立即过滤的水，在黑暗中冷藏的水样品的检测或eDNA浓度没有明显降低。暴露于光线和环境空气温度下的水样品从1小时开始未检出，到6 h这些样品中的大多数都低于检出限（LOD），到24 h在这些样品中均未检测到eDNA。这些结果对于无法立即获得冷藏的eDNA研究具有重要意义，或需要延长采样时间的野外工作（例如，划独木舟）。此外，我们报告了在环境条件下eDNA降解时间比以前的一些水族馆或中观宇宙研究更快，这表明研究与eDNA持久性和样品存储有关的机制的持续需求。