This study tested whether chords that do not differ in acoustic roughness but that have distinct affective connotations are strong enough to prime negative and positive associations measurable with an affective priming method. We tested whether musically dissonant chords low in valence (diminished, augmented) but that contain little acoustic roughness are strong enough in terms of negative affective connotations to elicit an automatic congruence effect in an affective priming setting, comparable to the major-positive/minor-negative distinction found in past studies. Three out of 4 hypotheses were supported by the empirical data obtained from four distinct sub-experiments (approximately N = 100 each) where the diminished and augmented chords created strong priming effects. Conversely, the minor chord and the suspended fourth failed to generate priming effects. The results demonstrate how automatic responses to consonant/dissonant chords can be driven by acquired, cultural concepts rather than exclusively by acoustic features. The obtained results of automatic responses are notably in line with previous data gathered from self-report studies in terms of the stimuli’s positive vs. negative valence. The results are discussed from the point of view of previous affective priming studies, cross-cultural research, as well as music historical observations.