Accumulating evidence suggests that assessment-informed personalized instruction, tailored to students’ individual skills and abilities, is more effective than more one-size-fits-all approaches. In this study, we evaluate the efficacy of Individualizing Student Instruction in Mathematics (ISI-Math) compared to Reading (ISI-Reading) where classrooms were randomly assigned to ISI-Math or ISI-Reading. The literature on child characteristics X instruction or skill X treatment interaction effects point to the complexities of tailoring instruction for individual students who present with constellations of skills. Second graders received mathematics instruction in small flexible learning groups based on their assessed learning needs. Results of the study (n = 32 teachers, 370 students) revealed significant treatment effects on standardized mathematics assessments. With effect sizes (d) of 0.41–0.60, we show that we can significantly improve 2nd graders’ mathematics achievement, including for children living in poverty, by using assessment data to individualize the mathematics instruction they receive. The instructional regime, ISI-Math, was implemented by regular classroom teachers and it led to about a 4-month achievement advantage on standardized mathematics tests when compared to students in control classrooms. These results were realized within one school year. Moreover, treatment effects were the same regardless of school-level poverty and students’ gender, initial mathematics or vocabulary scores.