Environmental DNA Marine Curacao 2020

Fish taxonomic, functional and phylogenetic composition variations recovered from eDNA metabarcoding along the coast of Curacao In February 2020, we collected a total of 20 water samples, from 10 stations, with two filtration replicates per station, in the two investigated coastal areas. Each station consisted of a transect of 2 km at an overall constant distance from the coast. We recorded the GPS coordinates at the start and end of the transect, which we used to map the transect positions. We conducted eDNA sampling by using a filtration device composed of an Athena® peristaltic pump (Proactive Environmental Products LLC, Bradenton, Florida, USA; nominal flow of 1.0 L/min), a VigiDNA® 0.20 μM cross-flow filtration capsule (SPYGEN, le Bourget du Lac, France) and disposable sterile tubing for each filtration capsule. We performed two filtration replicates in parallel on each side of a boat, at each station, for 30 min corresponding to a volume of ~30 L of water filtered by each capsule. At the end of each filtration, the water inside the capsules was emptied, and we filled the capsules with 80 ml of CL1 Conservation buffer (SPYGEN, le Bourget du Lac, France) and stored at room temperature. We followed a strict contamination control protocol in both field and laboratory stages (Valentini et al., 2016). Each water sample processing included the use of disposable gloves and single-use filtration equipment to avoid any risk of contamination. Libraries were prepared with ligation using the MetaFast protocol (Fasteris).库拉索海岸环境DNA宏条形码（eDNA metabarcoding）解析的鱼类分类学、功能学与系统发育组成变异 2020年2月，我们于两处调查沿岸区域的10个采样站位共采集20份水样，每个站位设置2组平行过滤重复样。每个站位的采样断面为离岸距离恒定的2千米沿岸样带。我们记录了样带起点与终点的GPS坐标，用于绘制采样断面位置。 我们采用由Athena®蠕动泵（Proactive Environmental Products LLC，美国佛罗里达州布雷登顿市；额定流量1.0 L/min）、VigiDNA® 0.20 μM错流过滤胶囊（SPYGEN，法国勒布尔热迪拉克市）以及适配每个过滤胶囊的一次性无菌管路组成的过滤装置开展环境DNA（eDNA）采样。每个站位在船舷两侧并行开展2组过滤重复实验，持续30分钟，对应每个过滤胶囊过滤约30 L水体。 每轮过滤结束后，排空胶囊内残留水体，随后向胶囊内注入80 mL CL1保存缓冲液（SPYGEN，法国勒布尔热迪拉克市），并置于室温保存。 我们在野外与实验室阶段均遵循严格的污染防控规程（Valentini et al., 2016）。每份水样的处理过程均使用一次性手套与一次性过滤设备，以规避任何污染风险。 采用Fasteris公司的MetaFast协议，通过连接法完成文库构建。