Water resource protection goals for aquatic life are often general and can result in under protection of some high quality water bodies and unattainable expectations for other water bodies. More refined aquatic life goals known as tiered aquatic life uses (TALUs) provide a framework to designate uses by setting protective goals for high quality water bodies and establishing attainable goals for water bodies altered by legally authorized legacy activities (e.g., channelization). Development of biological criteria or biocriteria typically requires identification of a set of least- or minimally-impacted reference sites that are used to establish a baseline from which goals are derived. Under a more refined system of stream types and aquatic life use goals, an adequate set of reference sites is needed to account for the natural variability of aquatic communities (e.g., landscape differences, thermal regime, and stream size). To develop sufficient datasets, Minnesota employed a reference condition approach in combination with an approach based on characterizing a stream's response to anthropogenic disturbance through development of a Biological Condition Gradient (BCG). These two approaches allowed for the creation of ecologically meaningful and consistent biocriteria within a more refined stream typology and solved issues related to small sample sizes and poor representation of minimally- or least-disturbed conditions for some stream types. Implementation of TALU biocriteria for Minnesota streams and rivers will result in consistent and protective goals that address fundamental differences among waters in terms of their potential for restoration.