Allowing students to practice science inquiry in the classroom is fundamental for science education. Students should master investigation skills, promoting their understanding of science concepts. Virtual laboratories have emerged as a powerful interactive tool that can be a suitable alternative to real laboratories, especially when lacking enough equipment and safety measures. This mixed approach study investigates the effects of virtual science laboratories on students’ motivation and attitude toward science (ATS). Students (n = 237) from grades (7, 8, 9, 10, and 11) were surveyed to evaluate their virtual laboratory experience in an American curriculum private school in Dubai with the theoretical basis of motivation theories. The study reported that virtual laboratories positively impact students’ overall motivation in terms of intrinsic motivation, perceived usefulness (U), effort (E), perceived self-efficacy (SE), and ATS. Students, perceived SE and U significantly impact student attitudes toward science (p < 0.001). Students’ perceived SE is a predictor of their E in the laboratory by 6.1%. Students with high SE put more E into completing a virtual laboratory task than students with low SE.