The soybean aphid (Aphis glycines) is an insect pest of cultivated soybeans (Glycine max). Several genes with resistance to A. glycines (i.e. Rag genes) have been identified in soybean. Virulent strains of soybean aphid are able to overcome the resistance and colonize soybeans having one or more Rag genes. It is important to classify virulent strains of soybean aphids in evaluating soybean lines in order to develop cultivars with durable resistance. The files presented here report the number of soybean aphids on soybean lines that differed in the Rag genes they contained. Two colonies of soybean aphid were tested. Materials and Methods Tests were conducted separately against the two soybean aphid colonies, which were maintained on soybean plants at North Central Agricultural Research Laboratory (NCARL), USDA-ARS, Brookings, South Dakota, USA, largely according to procedures described in Hesler and Tilmon (2018). The first colony was established from a single aphid collected near Volga, South Dakota, USA in 2016 and designated as ‘Volga16’ (Conzemius et al. 2019). It was reared on soybean cultivar ‘LD12R12-15805Ra’ (Rag1+Rag2 pyramid; University of Illinois, Urbana-Champaign, IL, USA). A second colony designated ‘Accrue’ was derived from a colony originally established from a single first instar isolated from aphids collected at Urbana, IL, USA, and initially reared in Urbana (‘Urbana clone’; Hill et al. 2004). This colony was established as an avirulent soybean aphid colony (Hill et al. 2004). A series of sequential colonies from the initial colony was established, in order, at The Ohio State University, Wooster, OH, USA; Iowa State University, Ames, IA, USA; South Dakota State University, Brookings, SD, USA; and finally, in 2018 at NCARL. Although established as an ostensibly avirulent colony derived from the ‘Urbana clone’ colony, it was unexpectedly virulent against a known resistant accession, LD05R-16137 (containing Rag1), in initial screening tests. Two separate no-choice tests were run for each soybean aphid colony. Each test consisted of seven soybean lines. Six had one or more Rag genes: 19APH18 (Rag1), 19APH25 (Rag2), 19INC (Rag3), 19APH29 (Rag4), 19APH30 (Rag6), 19APH09Rag12 (a Rag1+Rag2 pyramid); and ‘Titan,’ an aphid-susceptible soybean cultivar (Diers et al. 1999). Two-week-old, unifoliate-stage soybean plants growing in plastic pots (6 cm top diameter, 4 cm bottom diameter, 5.7 cm height) were each infested with 10 apterous adult soybean aphids and covered with a clear plastic, ventilated, cylindrical tube. After 20 days in an environmental chamber, the shoots of test plants were clipped at soil level, placed individually in sealable plastic bags, and stored in a freezer. Plants were removed over the next few days, and the aphids on them were counted. The data are contained in separate files—one for each of two soybean aphid colonies.