Despite the disproportional impact of HIV, Black individuals are benefiting the least from pre-exposure prophylaxis (PrEP). Motivational interviewing (MI) for PrEP uptake (MI-PrEP) is a two-session culturally tailored intervention incorporating MI strategies to improve PrEP motivation and uptake among cisgender Black women. A pilot randomized control trial was conducted in the Southeastern United States, and 41 women were randomized to MI-PrEP (session 1 with PrEP psychoeducation and MI and session 2 with MI and light case management) or enhanced treatment as usual (ETAU; two sessions of PrEP psychoeducation [videos explaining PrEP]). Women completed one follow-up assessment (1 month after visit 2). Measures captured primary (motivation [via contemplation and readiness ruler] and PrEP uptake via medical records) and secondary outcomes (e.g., PrEP knowledge, barriers to PrEP, and speaking to a provider about PrEP). Difference-in-differences analyses comparing MI-PrEP with ETAU as well as t-tests for within-group changes over time were conducted. Women who completed MI-PrEP (90.5% retained) compared with ETAU (100% retained) had a significantly higher likelihood of speaking to a provider about PrEP (OR = 4.42e7, CI [8.55e6, 2.29e8], DiD = 17.60, se = 0.84, p < 0.001). Within the MI-PrEP group, women had significant increases in PrEP prescription, knowledge, and motivation/contemplation, and significant decreases in financial resources as a PrEP barrier and medical mistrust (MMT). ETAU had within-group increases in PrEP prescription and speaking to a provider, no changes in motivation and MMT, and increases in specific barriers to care (e.g., transportation). MI-PrEP shows promise, and a large-scale study may be beneficial to further assess efficacy and examine implementation.