Sheffield United's first Johnstone's Paint Trophy campaign in more than 20 years continues with a trip to local rivals Rotherham in the Northern Section East.
The Blades overcame Burton in the first round 2-1 thanks to a last-gasp winner from Erik Tonne, and will now face further League Two opposition in the form of Andy Scott's Millers.
Preston are back in the competition after relegation from the Championship and will visit Morecambe, who also received a bye to the second round, in the Northern Section West.
Crewe take on Macclesfield after dispatching League One side Bury on penalties, while Walsall travel to Rochdale after Ryan Jarvis' wonder goal saw the Saddlers squeeze past Shrewsbury.
In another Yorkshire derby, Bradford travel to the Galpharm Stadium to take on Huddersfield in the Northern Section East. The Bantams dismissed Sheffield Wednesday in a penalty shootout at Valley Parade, with the Terriers taking a 2-1 victory from Northampton at Sixfields.
Oldham received a bye to the second round and travel to Scunthorpe. Paul Dickov's Latics take on an Iron side who dispatched Hartlepool 2-0 at Glanford Park in the previous round.
League Two new boys AFC Wimbledon drew Stevenage in the Southern Section East courtesy of both receiving byes in the opening round. Charlton take on last year's beaten finalists Brentford and Gillingham welcome Barnet, who stunned Colchester at Whaddon Road.
Bournemouth take on Yeovil after demolishing Hereford 4-1 in the first round and Aldershot draw Chris Wilder's Oxford after both sides received byes to this stage.
Exeter will play Swindon after reaching the second round of the Southern Section West following a penalty shootout victory over fierce Devon rivals Plymouth. Accrington or Carlisle will be at home to Tranmere with Dagenham or Leyton Orient facing Southend at home.
Notts County will host Chesterfield and the winner of Tuesday night's clash between Wycombe and Bristol Rovers will face Cheltenham.
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