The regiment was formed as the 127th Ohio Infantry Regiment at Camp Delaware, Ohio. It was re-designated the 5th United States Colored Infantry Regiment, and moved to Norfolk, Virginia, in November 1863, immediately after three months of organization. It served at Norfolk and Portsmouth in the Department of Virginia and North Carolina until January 1864, during which time the unit participated in Brigadier GeneralEdward A. Wild's expedition to South Mills and Camden Court House, North Carolina, from December 5 to December 24 and in action at Sandy Swamp, North Carolina, on December 8.
General Benjamin F. Butler remarked "[s]o far as the conduct of the color-sergeant, (Milton M.) Holland, was concerned, in the charge at New Market Heights, had it been within my power I would have conferred upon him in view of it, a brigadier-generalship for gallantry on the field."[1]
In December the unit was assigned to the newly formed XXV Corps and took part in the failed attack on Fort Fisher, North Carolina, from December 7 to December 27, 1864, and the successful Second Battle of Fort Fisher from January 7 to January 15, 1865. The 5th then saw action at Sugar Loaf Hill on January 19 and at Federal Point on February 11 before taking part in the Battle of Wilmington at Fort Anderson from February 18 to February 20 and the capture of Wilmington as well as action at Northeast Ferry on February 22, 1865.
In March 1865, the regiment was reassigned to the X Corps and took part in General William Tecumseh Sherman's Carolinas Campaign. The unit saw action during the advance on Kinston and Goldsboro, North Carolina, starting on March 6 and occupied Goldsboro after its capture on March 21. The regiment saw further action at Cox's Bridge on March 23 and March 24 and participated in the advance on Raleigh, North Carolina, starting on April 10 and the occupation of Raleigh after the city's fall on April 13. With the end of the war at hand, the men of the regiment witnessed the surrender of Confederate General Joseph E. Johnston and his army at Bennett Place, North Carolina, on April 26, 1865. The unit served out the rest of its term in Goldsboro, New Bern, and Carolina City, North Carolina.
The 5th United States Colored Infantry was mustered out on September 20, 1865, after two years of existence. The regiment lost a total of 249 men during its service; four officers and 77 enlisted men were killed or mortally wounded and two officers and 166 enlisted men died of disease.