These data were collected in support of the sampling goals of the Aquatic Intermittency effects on Microbiomes in Streams (AIMS) Project. We sampled macroinvertebrates along a 100 m reach every 20 m alternating channel location from left, to center, to right for a total of six samples per reach. We compiled all six samples into a composite sample, removed debris and leaf matter via manual inspection of material and elutriation, and sieved the remaining sample through a 500 µm sieve. We then preserved the sample in 95% ethanol and returned it to the lab. In the lab, ethanol was refreshed before we froze the sample in a -20 ºC freezer. We shipped samples to a commercial lab (Jonah Ventures, Boulder, CO) for metabarcoding analysis of the invertebrate community. Briefly, Jonah Ventures homogenized community samples with a hand immersion blender and extracted DNA with a DNeasy Powersoil Kit following the manufacturer’s protocol. They then amplified samples using two primers from the CO1 gene (CO1 F230 fragment Hajibabaei et al. 2012; CO1 BE fragment Folmer et al. 1994, Gibson et al. 2015). They conducted PCRs with initial denaturation at 95 ºC for 5 min, followed by 40 cycles of 40s at 95 ºC, 1 min at 46 ºC, 30s at 72 ºC and a final elongation at 72 ºC for 10 min. They then cleaned samples using Exo1/SAP, and pooled, normalized and indexed. Samples were sent for sequencing at the CU Boulder BioFrontiers Sequencing Center where the Center used the v2 500-cycle kit with appropriate quality control measures. Jonah Ventures then demuliplexed sequenced samples using phigs v2.1.0 followed by removal of gene primers and merging read pairs. Read pairs were then clustered using unoise3 denoising algorithm in vsearch and with sequences with less than 8 reads discarded. Taxonomy was assigned using a custom best-hits algorithm with reference to NCBI Gen Bank to each Exact Sequence Variant (ESV) (See linked Jonah Venture documents for full methods).

JDR - Johnston Draw is a 1.8-km2 watershed in southwestern Idaho, USA (outlet location: 43.1226, -116.776) located within the Reynolds Creek Critical Zone Observatory in western Idaho, a research center with cattle grazing. Elevation ranges from approximately 1490m to 1850m. The mean annual precipitation in the watershed is 550 mm/yr with rainfall occurring at the lower elevations and snowfall, resulting in large drifts, at the higher elevations (Godsey et al., 2018). Mean annual temperatures range from 8.9C at the bottom of watershed and 4.7C near the top.
GBJ - Located near Pocatello, ID, Gibson Jack Creek (outlet location: 42.7853, -112.4446) drains 1620 ha of the US Forest Service Research Natural Area within the Caribou National Forest. Predominantly forested with deciduous trees, sub-apline fir, Douglas fir, and with woody shrubs, sagebrush, and grasses, Gibson Jack spans an elevation range of 1555-2130 m, and has an mean annual temperature and precipitation of 6.5°C and 614.5 mm/yr, respectively. Gibson Jack spans the rain to snow transition with rainfall occurring at the lower elevations and snowfall at the upper elevations. Gibson Jack Creek drains to the Portnuef River and is heavily recreated by the local community.
DRC - Con1W in Dry Creek Experimental Watershed (outlet location: 43.730136, -116.134807), run by Boise State University and located near Boise, ID, is predominantly forested by Ponderosa Pine and Douglas fir, with woody shrubs, sagebrush, and grasses. With elevations from 1295-1907 m, Con1W in Dry Creek spans the rain-snow transition with rainfall occurring at lower elevations and snowfall at the upper elevations in the semi-arid southwestern region of Idaho. Con1W in Dry Creek has mean annual temperatures of 8.3°C and mean annual precipitation at 648.8 mm/yr.