Soybean aphid, Aphis glycines Matsumura, a pest of soybean, Glycine max (L.) Merr., and native of Asia, invaded North America sometime before 2000 and rapidly became the most significant insect pest of soybean in the upper Midwest. Plant resistance, a key component of integrated pest management, has received significant attention in the past decade, and several resistance (Rag) genes have been identified. However, the efficacy of Rag (Resistance to Aphis glycines) genes in suppressing aphid abundance has been challenged by the occurrence of soybean aphids capable of overcoming Rag gene-mediated resistance. Although the occurrence of these Rag virulent biotypes poses a serious threat to effective and sustainable management of soybean aphid, little is known about the current abundance of biotypes in North America. The objective of this research was to determine the distribution of Rag virulent soybean aphids in Wisconsin. Soybean aphids were collected from Wisconsin during the summers of 2012 and 2013, and assayed for Rag1, Rag2, and Rag1 2 virulence using no-choice tests in a greenhouse. One clone from Monroe County in 2012 reacted like biotype 4, three clones in different counties in 2013 responded like biotype 2, and eight others expressed varying degrees of Rag virulence. Rag virulence in 2013 was observed in aphids from 33% of the sampled sites and was accounted for by just 4.5% of sampled clones, although this is likely a conservative estimate. No-choice test results are discussed in light of current questions on the biology, ecology, and population genetics of soybean aphid.