Two second-half goals from visitors Yeovil denied Reading top spot in Southern group A, as Glovers keeper John Maddison put on a fine display for the away side in goal.
Reading were undone by free kicks on the edge of their area on two occasions, when first Tom Eaves and then Francois Zoko found the net from close range.
A Reading side that were dominant in possession were denied time and again by the Yeovil stopper, who put on an impressive display to keep out efforts from Josh Barrett, Niall Keown, Sam Smith and Jake Cooper with several fine saves.
Martin Kuhl named a young starting XI for the Checkatrade Trophy clash, with ten of the starting line up coming through the Reading youth system.
Dominic Samuel who was named captain, Liam Kelly, Tennai Watson and Jake Cooper were all named to start for the Royals, with the latter continuing his return to fitness following a lengthy injury lay-off.
Kuhl’s young Reading side started the contest on the front foot, with Andrija Novakovich causing all kinds of problems for the Yeovil defence down the right flank.
In a first half of few chances Reading enjoyed the majority of possession. Jake Cooper went close for Reading after 20 minutes, when he rose highest to meet a Liam Kelly free kick. The giant defender was only able to steer his header at Yeovil keeper John Maddison from six yards out.
Yeovil’s best effort came through midfielder Matt Dolan, who found half a yard of space on the edge of the Reading box before unleashing a low drive that was well saved by young Reading stopper Stewart Moore.
Glovers’ right-back Liam Shephard shot just wide for the visitor’s when he drove up the right flank before shooting from a tight angle.
Towards the end of the first period Liam Kelly went close for Reading when his cross-shot fizzed past of the Yeovil upright with Maddison scrambling across his line.
A couple of minutes into the second half the visitors had the lead. A deep free kick from Yeovil saw half-time substitute Tom Eaves breach the Royals offside trap and finding himself one-on-one with Stuart Moore. The striker made no mistake from inside the Reading box, finishing low into the bottom corner to hand the Glovers the lead.
Reading looked to respond, with Tyler Frost and Andy Rinomhota both having efforts fly over the opposition crossbar in the following minutes.
Substitute Josh Barrett had Reading’s best opportunity of the night until that point when his low shot from inside the area was saved brilliantly by Maddison. Great work on the right wing from Tannai Watson picked out the midfielder, but Barrett was unable to convert.
Barrett was denied again five minutes later after a powerful run down the right flank. The Republic of Ireland U19 international made his way into the opposition box before hammering an effort at goal, forcing a smart save from Maddison.
As the half progressed it looked as though Reading would find a way back into the game but for more heroics from Maddison. The Yeovil keeper produced a string of saves in the last 20 minutes to keep the Royals at bay.
Sam Smith went the closest for Reading when he saw his flicked effort kept out by the glovesman. Novakovich’s low shot was diverted by the substitute but Maddison pulled off a great reaction stop to deny him. From the resulting corner the keeper was again called into action when Niall Keown rose highest to head at goal, but the keeper was there again.
Despite the late flurry of Reading pressure it was Yeovil that managed to find the net for the second time when they hit the Royals on the counter attack. The Glovers won a free-kick on the edge of the Reading area, with the set piece deflected through the wall falling kindly for substitute Francois Zoko. The forward slotted home from close range to seal the match for Yeovil and send them through as group winners.
More outstanding keeping from Maddison late on stopped a Reading consolation, when the keeper pulled off a fantastic double save to deny Jake Cooper from close range.
Full time: Reading 0-2 Yeovil Town
Reading: Moore, Watson, Cooper, McIntyre, Richards (Smith 79'), Kelly (Barrett 45'), Gravenberch, Rinomhota, Frost, Novakovich, Samuel (Keown 63').
Unused subs: Tupper, Andresson, East, Vancooten.
Yeovil Town: Maddison, Shephard, Smith, Dolan, Mugabi, Lacey, Khan (Zoko 70'), Butcher (Ezewelle 81'), Campbell (Eaves 45'), Hedges, Whitfield.
Unused subs: Krysiak, Copp, Sowunmi.