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Background Early diagnosis is crucial to reduce the morbidity and mortality associated with breast cancer (BC). Awareness of BC symptoms plays a key role in this. This study aimed to evaluate the Palestinian women’s awareness of BC symptoms and determine factors associated with good awareness. Methods This was a national cross-sectional study conducted from July 2019 to March 2020 in Palestine. Convenience sampling was used to recruit adult women from hospitals, primary healthcare centers, and public spaces located in 11 governorates. A translated-into-Arabic version of the validated BC awareness measure was utilized for data collection. The awareness level was categorized based on the number of symptoms recognized into: poor (0 to 4), fair (5 to 9), and good (10 to 13). Results Of 6269 approached, 5434 participants completed the questionnaire (response rate = 86.7%). A total of 5257 questionnaires were included in the analysis: 2551 from the Gaza Strip and 2706 from the West Bank and Jerusalem (WBJ). Participants living in the WBJ were more likely to be older, have higher monthly income, and suffer from more chronic diseases than participants living in the Gaza Strip. The most frequently identified BC symptom was ‘lump or thickening in the breast’ (n = 4887, 92.9%) followed by ‘lump or thickening under the armpit’ (n = 4394, 83.6%). The least frequently identified symptoms were ‘pulling in of the nipple’ (n = 2665, 50.7%) and ‘change in the position of the nipple’ (n = 2710, 51.6%). A total of 2191 participants (41.7%) demonstrated good awareness of BC symptoms. Participants from the Gaza Strip were more likely than participants from the WBJ to have good awareness (47.0.0% vs. 36.7%). On the multivariable analysis, being ≥ 40 years, completing a post-secondary education, knowing someone with cancer, and visiting hospitals and primary healthcare centers were all associated with an increase in the likelihood of having good awareness. However, living in the WBJ was associated with a decrease in the likelihood of having good awareness. Conclusion Less than half of women included in this study showed good awareness of BC symptoms. More targeted educational interventions are needed to promote Palestinian women’s awareness of BC symptoms to facilitate early diagnosis.