Our previous functional near-infrared spectroscopy (fNIRS) study on motor sequence learning (Polskaia et al., 2020) did not detect the same decrease in activity in the left dorsolateral prefrontal cortex (DLPFC) associated with movement automaticity, as reported by Wu et al. (2004). This was partly attributed to insufficient practice time to reach neural efficiency. Therefore, we sought to expand on our previous work to better understand the contribution of the prefrontal cortex (PFC) to motor sequence learning by examining learning across a longer period of time. Participants were randomly assigned to one of two groups: control or trained. fNIRS was acquired at three time points: pre-test, post-test, and retention. Participants performed four sequences (S1, S2, S3, and S4) of right-hand finger tapping. The trained group also underwent four days of practice of S1 and S2. No group differences in the left DLPFC and ventrolateral (VLPFC) were found between sessions for S1 and S2. Our findings revealed increased contribution from the right VLPFC in post-test for the trained group, which may reflect the active retrieval of explicit information from long-term memory. Our results suggest that despite additional practice time, explicit motor sequence learning requires the continued involvement of the PFC