Gary Smith's second-half header gave Darlington a hard-fought second away victory of the season.
The U's missed a golden chance to take the lead from the penalty spot after Aman Verma fouled Luke Berry, but Berry's shot was saved by Sam Russell.
The visitors made Cambridge pay, with Smith rising to head home a left-wing cross from Aaron Brown.
The hosts had the chance to level the match late on but Danny Wright volleyed over the crossbar.
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Cambridge Utd:
Brown, Roberts, Coulson, Saah, Coakley, Patrick, McAuley (Marriott 54), Berry (Platt 72), Wellard, Hughes, Wright. Subs Not Used: Naisbitt, Ives, Brighton.
Darlington:
Russell, Arnison, Miller, Burn, Brown, Verma, Taylor, Gary Smith, Bridge-Wilkinson, Hatch, Modest (Michael Smith 90). Subs Not Used: St Louis-Hamilton, Senior, McReady, Quinn.
Booked: Hatch, Verma.
Goals: Gary Smith 50.
Att: 1,635
Ref: Chris Knowles (Northamptonshire).
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