Background: Breast cancer imposes a significant global health burden not only due to being constantly ranked as the leading malignancy in females, but also due to the immense resulting morbidity and mortality. Despite the presence of screening modalities, the uptake is relatively low due to unidentified barriers in many countries and regions. The Identification of these barriers towards screening is of paramount importance to tackle the health burden imposed by breast cancer, especially in countries like Jordan, a developing middle eastern country. Methods: A self-reporting survey consisting of comprehensive questions regarding the participant’s demographics, medical history, knowledge about breast cancer and the screening modalities was distributed to females aged 25 years and above with no prior history of breast cancer who are residing across the different regions of Jordan. The first consecutive 790 participants were enrolled, and data analysis was performed to identify the key barriers. Results: The mean age of the enrolled study population was 41 ± 9 years old with the majority being married (82.8%), have underwent a higher level of education (86.5%), are non-smokers (75.2%), and are medically insured (72.4%). 209 participants (26.5%) had reported a family history of breast cancer, while 83.4% of the participants reported awareness about breast cancer screening modalities with breast self-examination being the most commonly known modality of screening in 39.2% of the participants. Only 22.4% of the participants were aware of the 3 available screening methods. The majority of the participants (57.1%) did not undergo previous screening. When the participants who did not undergo previous breast cancer screening were compared with those who did, the participants without prior screening were more likely to report the lack of knowledge towards screening modalities (27.7% vs 8.0%) (p-value < 0.001), fear of the results (22.6% vs 13.9%) (p-value = 0.002), fear of pain during or after from the test (10.4% vs 5.9%) (p-value = 0.024), and feeling of embarrassment provoked by the screening test (15.7% vs 6.5%) (p-value < 0.001) as key barriers. On the contrary, there was no statistically significant difference between the two groups when assessing financial aspect of screening uptake as a potential barrier (p-value = 0.860). Conclusions: Despite the considerably high percentage of participants being aware of breast cancer screening modalities, an alarmingly low rate of screening uptake was witnessed due to several barriers. The collective identification of lack of knowledge towards screening modalities and the nature of the available tests, in addition to the fear of the results and pain following the test, and feeling of embarrassment undoubtedly mandates the formulation of comprehensive awareness campaigns that effectively tackle these barriers.