The young Millers fell tasted just their second defeat of the Youth Alliance campaign on Friday afternoon as they were narrowly beaten by Burton Albion at Sheffield Hallam University Sports Park.
Jerome Greaves had given Rotherham an advantage with half an hour played, but the away side responded to head home from a set piece to level proceedings.
In-form Greaves did find the net once more midway through the second half, although it meant nothing in the end as Albion rallied to secure all three points in a 3-2 victory.
Millers | Ellis; Millen, Makwedza (Salah 73'), Hull, Smith; Peel-Evans, Spence (C), Gratton, Hodgkinson (Gateshill 59'); Kenny; Greaves (Scott 80').
Unused | Wenham.