This study aimed to identify the differentiated instruction practices used by Jordanian teachers and the challenges they faced when teaching students with learning disabilities in Amman. The sample of the study consisted of 194 teachers. It followed a mixed method design and consisted of two parts. First, a quantitative analysis of a questionnaire of 75 items was developed on six domains (differentiation in content, process, resources, product, assessment, and differentiation in learning environment). Second, a qualitative analysis of interviews with teachers was conducted. Validity and reliability were established. Results of the study showed that the mean of the scores for the six domains and the entire items are low. ANOVA analysis showed that there were no statistically significant differences related to the variable of teacher's experience. On the other hand, there were statistically significant differences, as to the type of school, in favor of private schools. The main challenges, as the study found out, were weak administrative support, low parental support, lack of time, and shortages in learning resources.