RELEASE-2023

Benthic microbe samples are collected at the same time and location as periphyton (microalgae) samples three times per year in wadeable streams during aquatic biology bout windows, roughly in spring, summer and fall. Samples are collected using aseptic methods using the most appropriate sampler for the habitat and substratum type, including rock scrubs, grab samples, and epiphyton. Samples are collected in the two most dominant benthic habitat types (e.g. riffles, runs, pools, step pools). Cobble scrubs are filtered on 0.22 um Sterivex capsule filters, capped and flash-frozen in the field. Grab samples of sediment (silt, sand) or plant material/epiphyton are collected when appropriate. All samples are frozen on dry ice in the field and are shipped frozen to an analytical facility for DNA sequence analysis using primer sets targeting the small subunit of the ribosomal RNA gene. Sequence data undergo QAQC and are processed bioinformatically using an in-house pipeline to identify unique sequence variants in the data and to taxonomically identify each sequence variant. Instrumentation:

本数据集的底栖微生物（benthic microbe）样本与周丛生物（periphyton，微藻）样本，于每年三次的可涉水溪流水生生物调查窗口期内，在相同的时间与地点采集，采样时段大致覆盖春季、夏季与秋季。样本采用无菌操作方法采集，将根据生境与底质类型选用最合适的采样器具，涵盖岩石刷洗采样、抓斗采样以及附生生物（epiphyton）采样等方式。采样将在两类最具代表性的底栖生境中开展，例如急流浅滩（riffles）、缓流区（runs）、深潭（pools）与阶梯式深潭（step pools）。砾石刷洗样本将通过0.22微米Sterivex胶囊式滤器进行过滤，封装后于采样现场完成快速冷冻。若条件适宜，将采集沉积物（粉砂、砂粒）或植物材料/附生生物的抓斗样本。所有样本均在采样现场以干冰冷冻保存，并以冷冻状态运至分析机构，采用靶向核糖体RNA基因小亚基的引物组开展DNA序列分析。序列数据将先经过质量保证与质量控制（QAQC）流程，随后通过自研生物信息学分析流程进行处理：一是识别数据中的独特序列变异体，二是对每一个序列变异体开展分类学鉴定。仪器设备：