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The National Ecological Observatory Network (NEON) seeks to facilitate ecological prediction at a continental scale by measuring processes that drive change and responses at sites across the United States for thirty years. The spatial distribution of observations of terrestrial organisms and soil within NEON sites is determined according to a "design-based" sample design that relies on the randomization of sampling locations. Development of the sample design was guided by high-level NEON objectives and the multitude of data products that will be subjected to numerous analytical approaches to address the causes and consequences of ecological change. A requirement framework permeates the NEON design, ensuring traceability from each facet of the design to the high-level requirements that make the NEON mission statement actionable. Requirements were developed for the terrestrial sample design to guide the key components of the design:

1. Randomizing the sample locations ensures the unbiased collection of data, is appropriate for organisms and soil, and provides data suitable for a variety of analyses.

2. Stratification increases efficiency and allows sampling to focus on those parts of the landscape measured by other NEON observation platforms.

3. Attention to the sample size and spatial plot allocation ensures that data products will be sufficient to inform questions asked of the data and the NEON objectives.

4. Establishing a framework with the capacity for re-evaluate and design iteration allows for adaption to unexpected challenges and optimization of the sample design based on early data returns.

The utility of the NEON sampling design is highlighted by its application across terrestrial systems. The data generated from this unique design will be used to quantify patterns in: the abundance and diversity of small mammals, breeding birds, insects, and soil microbes; vegetation structure, biomass, productivity, and diversity; and soil biogeochemistry.