Purpose: The purpose of the study was to develop Evidence-based nursing practice guideline for post-lumbar puncture patients in neurology ward and to evaluate the guidelines by applying them to practice. Methods: Eighty patients without preceding headache who underwent Lumbar puncture were selected prospectively. We compared the incidence of post-lumbar puncture headache between bed-rest for 4 hours and 1hours. Results: The post-lumbar puncture headache was found in 13 cases among bed-rest for 4 hours and 1hours groups. The frequency of each group was 25%(10/40) in 4 hours-bed rest group and 7.5%(3/40) in 1 hours-bed rest group. Conclusion: There was no significant difference between two groups (p-value= .066). 4 hours-bed rest after lumbar puncture is unnecessary.