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Ashlee Barnes has a strong background in nursing, with a range of roles and responsibilities. Ashlee started their career at the University of Iowa Hospitals and Clinics as a Registered Nurse, where they provided care to surgical patients and administered medication. Ashlee also gained experience in wound care. In 2009, they joined Great River Health as a Registered Nurse, specializing in labor and delivery. Ashlee provided total care to women during pregnancy, labor, postpartum, and for newborn and NICU patients. In 2018, they transitioned to a role as a Clinical Facilitator, and later in 2019, they became a Nursing Manager. Currently, they serve as the Director of Nursing at Great River Health.

Ashlee Barnes completed their Associate's Degree in Nursing, with a focus on Nursing Education, at Southeastern Community College from 2005 to 2008. Ashlee later pursued a Bachelor's degree in Nursing at Western Governors University. Additionally, they hold certifications in Basic Cardiac Life Support, but the institution and dates of completion are unknown. Ashlee also has certifications in Neonatal Resuscitation Program and Pediatric Advanced Life Support, but the institutions and dates are unspecified.