The so-called control-of-variables strategy (CVS) incorporates the important scientific reasoning skills of designing controlled experiments and interpreting experimental outcomes. As CVS is a prominent component of science standards appropriate assessment instruments are required to measure these scientific reasoning skills and to evaluate the impact of instruction on CVS development. A detailed review of existing CVS instruments suggests that they utilize different, and only a few of the four, critical CVS sub-skills in the item development. This study presents a new CVS assessment instrument (CVS Inventory, CVSI) and investigates the validity of student measures derived from this instrument utilizing Rasch analyses. The results indicate that the CVSI produces reliable and valid student measures with regard to CVS. Furthermore, the results show that the item difficulty depends on the CVS sub-skills utilized in item development, but not on the item content. Accordingly, previous instruments that are restricted to a few CVS sub-skills tend to over- or underestimate students’ CVS skills. In addition, these results indicate that students are able to use CVS as a domain general strategy in multiple content areas. Consequences for science instruction and assessment are discussed.