Background: Hamstrings undergo adaptive shortening. It is common in healthy individuals to have a sedentary lifestyle affecting males (91.8%) more than females (78.7%). Hamstring tightness can interrupt ADL’s predisposing the person to further health problems i.e. plantar fasciitis, sacroiliac joint pain, decreased lumbar lordosis, knee pain, quadriceps malfunction, postural disturbances, overstretched lumbar ligaments, reduced muscle strength. Aims and Objectives: To analyze the prevalence of hamstring tightness and its effects on lower extremity function in individuals with prolonged standing. Materials and Methods: 65 participants were chosen for this cross-sectional study by convenient sampling; all were female nurses. The hamstring tightness was measured by the data collection tools i.e. lower limb task questionnaire, straight leg raise test, and active knee extension test after having the consent forms signed by the participants. The data was collected from three big hospitals in the city. Results: The results of the study conducted to see the prevalence of hamstrings tightness and its impact on lower extremity function in asymptomatic individuals with prolonged standing hours showed that only 15.38% of the sample (n=65) with a mean age of 31-35 years were found with bilateral hamstrings tightness while 84.62% were found with no hamstrings tightness. Conclusion: The prevalence of hamstring tightness in asymptomatic individuals with prolonged standing hours is low.