Recent studies of eurypterid paleoecology suggest that formation of eurypterid Lagerstätten in the mid-Paleozoic of Laurentia was controlled by the presence of an ecological–taphonomic window that recurred predictably in nearshore, marginal environments during transgressions. We tested this hypothesis by performing a high-resolution taxonomic, environmental, and stratigraphic survey and quantitative analysis of all Silurian–Lower Devonian eurypterid-bearing intervals in the Appalachian basin, the most prolific region for eurypterid remains in the world. Canonical correspondence analysis of sedimentological and faunal associations revealed a strong lithologic gradient between groupings of eurypterid genera and associated taxa across the basin, and a significant association of eurypterids with microbialites (thrombolites, stromatolites) and evaporitic structures. Field observations confirmed that, stratigraphically, eurypterids in the basin frequently occur above the microbialite structures and beneath evaporites and other indicators of increased salinity or subaerial exposure. Following interpretation of these features within a sequence stratigraphic framework, we present a preservational model in which (1) eurypterids inhabited nearshore settings following freshening conditions concomitant with minor transgressions, (2) their remains were subsequently buried by storms or microbialite sediment baffling, and (3) subsequent long-term preservation of tissues was facilitated by regression and cyclical shallowing-up successions that promoted hypersalinity and anoxia. In the central and southern region of the basin, where microbial structures and evidence for hypersalinity are less common, a similar pattern of cyclical shallowing-upward deposition within eurypterid-bearing units holds. Thus, eurypterid preservation appears to reflect a combination of ecological preferences and abiotic conditions that promoted inhabitation and eventual preservation within the same setting. This study provides the first quantitative support for a sea level–based control on preservation of eurypterids and adds to the growing body of evidence that suggests that analysis of exceptional preservation in the fossil record benefits from interpretation within a sequence stratigraphic framework.