Gillingham provided the biggest shock of the Carabao Cup first round by knocking out last season’s semi-finalists Southampton.
Ashley Nadesan’s goal and a Robbie McKenzie double saw the League Two side beat the team just relegated from the Premier League 3-1 at Priestfield.
Saints boss Russell Martin made 11 changes from the side that won at Sheffield Wednesday on Friday, but it backfired spectacularly despite a late consolation from substitute Carlos Alcaraz.
Carlos Alcaraz scored a late consolation goal for Southampton
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Highlights of the first-round Carabao Cup clash between Gillingham and Southampton
Hull were also Championship casualties, beaten 2-1 by League Two Doncaster thanks to two goals from George Miller after Oscar Estupinan had given the Tigers the lead.
Millwall joined them through the exit door after two goals from Kelvin Ehibhatiomhan and further strikes from Charlie Savage and Mamadi Camara gave troubled Reading a thumping 4-0 win at The Den.
Three Championship sides were dumped out on penalties by lower-division clubs. Sunderland lost the shootout 5-3 to Crewe after a 1-1 draw, Stevenage held their nerve to knock out Watford and Salford took the scalp of Preston.
Sheffield Wednesday needed a Tyreeq Bakinson equaliser seven minutes into stoppage time to earn a 1-1 draw with Stockport, with the Owls winning the penalty shootout 4-1.
Aiden Marsh left it even later, equalising for Barnsley in the 98th minute to earn a 2-2 draw with Tranmere, but League Two Rovers triumphed 7-6 on spot-kicks.
League Two Newport fought back from a goal down to beat League One Charlton 3-1.
Newly-promoted Wrexham, playing their first League Cup tie since 2007, went through on penalties after a goalless draw at home against Wigan.
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Highlights of the first-round Carabao Cup clash between Wrexham and Wigan
League Two side Wrexham made it past Wigan on penalties
Peterborough won on spot-kicks after a 1-1 draw with Swindon, as did Rotherham, who drew 1-1 at home to Morecambe.
Sutton were also victorious from 12 yards after a 2-2 draw with Cambridge, and Bradford saw off League Two rivals Accrington 4-1 on penalties following a 1-1 draw.
Stoke beat West Brom 2-1 and Middlesbrough won the other all-Championship clash at Huddersfield 3-2.
Joel Piroe struck twice in Swansea’s 3-0 home win over Northampton to give new boss Michael Duff his first victory, and two goals from Juninho Bacuna saw Birmingham through with a 2-0 success at Cheltenham.
There were also wins for Blackpool, Bolton, Plymouth, Port Vale, Portsmouth, Exeter, Mansfield, Lincoln, Wycombe and Harrogate.