Rovers, who were top of Group E at kick-off, conceded four first-half goals and gave away a penalty as they were embarrassed by what was believed to be Barnsley's youngest-ever side in the opening period.
Sixteen-year-old Fabio Jalo helped himself to two goals for the visitors along with former Doncaster loanee Josh Martin.
A second-half brace from substitute George Miller helped salvage some pride, however, with the game finishing 4-2 to Barnsley, who secured qualification.
Doncaster now need Newcastle’s under-21s to take points from Lincoln to qualify.
![George Millers takes a shot at goal.](https://www.doncasterfreepress.co.uk/webimg/b25lY21zOmRjZGJhZmYxLTJjZGEtNGY3Yi05NTM1LTg0MmUxMGM4ZmZhZjoxNjViYTA0NS01ZmI3LTQ5MWYtOGQxMi1lNzFlNjc4NzE5YzY=.jpg?crop=3:2&width=640)
![Left with little chance given some pathetic defending for the goals but emerged with some credibility after saving a penalty then denying several more goalbound efforts.](https://www.doncasterfreepress.co.uk/webimg/b25lY21zOjhiYWY1NTYyLTBiYjAtNDcyZS04Njk2LWU4Mzc3NWZlZWMwNTpiZDBhODZhOC1kODNjLTQwYmYtOGM3OC00ZjU4NWViMmI0YjM=.jpg?crop=3:2&width=640)
2. Louis Jones - 6
Left with little chance given some pathetic defending for the goals but emerged with some credibility after saving a penalty then denying several more goalbound efforts. Photo: Howard Roe/ahpix.com
![Responsible for the first goal when his missed clearance allowed Slobodan Tedic through on goal, who passed to Josh Martin for a tap-in. Later brought Tedic down for a penalty.](https://www.doncasterfreepress.co.uk/webimg/b25lY21zOmNiZTViOThlLWFhMjUtNDgwMC05N2VmLTc4NWJhNjk5MWNlODowZjgzNDdjZS0zNWRmLTRiYTktYjk4OS04MzE3ZDQ2OWU5MTc=.jpg?crop=3:2&width=640)
3. Tom Anderson - 4
Responsible for the first goal when his missed clearance allowed Slobodan Tedic through on goal, who passed to Josh Martin for a tap-in. Later brought Tedic down for a penalty. Photo: Howard Roe/AHPIX LTD
![Not directly responsible for any goal but can't be exonerated from a dreadful first-half defensive display.](https://www.doncasterfreepress.co.uk/webimg/b25lY21zOjUxZDUwYWM4LTU5MmMtNGEwYy05NWFkLTFiM2ZhODA3NGM3NDo3MWU5NDM4Yy1lN2Q0LTQyZTMtYWQ3My0xMjEyZGQ4NjY3NmI=.jpg?crop=3:2&width=640)
4. Adam Long - 5
Not directly responsible for any goal but can't be exonerated from a dreadful first-half defensive display. Photo: Howard Roe/ahpix.com