Current pain assessment and management in neonates need to be fully described before neonatal pain care can be optimized. This study’s purpose was to report neonatal nurses’ knowledge, existing pain assessment practice, and pharmacological pain management of neonates in Jordan. A cross-sectional descriptive study was conducted. Eighteen neonatal intensive care units in Jordan were included in the study. One hundred eighty-four neonatal nurses participated. Questionnaires were distributed by and returned to the neonatal intensive care units’ managers between June and August 2014.