Listeners are adept at extracting regularities from the environment, a process known as statistical learning (SL). SL has been generally assumed to be a form of “context-free” learning that occurs independently of prior knowledge, and SL experiments typically involve exposing participants to presumed novel regularities, such as repeating nonsense words. However, recent work has called this assumption into question, demonstrating that learners’ previous language experience can considerably influence SL performance. In the present experiment, we tested whether previous knowledge also shapes SL in a non-linguistic domain, using a paradigm that involves extracting regularities over tone sequences. Participants learned novel tone sequences, which consisted of pitch intervals not typically found in Western music. For one group of participants, the tone sequences used artificial, computerized instrument sounds. For the other group, the same tone sequences used familiar instrument sounds (piano or violin). Knowledge of the statistical regularities was assessed using both trained sounds (measuring specific learning) and sounds that differed in pitch range and/or instrument (measuring transfer learning). In a follow-up experiment, two additional testing sessions were administered to gauge retention of learning (one day and approximately one-week post-training). Compared to artificial instruments, training on sequences played by familiar instruments resulted in reduced correlations among test items, reflecting more idiosyncratic performance. Across all three testing sessions, learning of novel regularities presented with familiar instruments was worse compared to unfamiliar instruments, suggesting that prior exposure to music produced by familiar instruments interfered with new sequence learning. Overall, these results demonstrate that real-world experience influences SL in a non-linguistic domain, supporting the view that SL involves the continuous updating of existing representations, rather than the establishment of entirely novel ones.