The A-road will be shut northbound between Hinksey Hill and Peartree Interchange from 9pm.
It is expected to reopen at 6am tomorrow (November 7) as National Highways resurfacing works are carried out.
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National Highways has also announced there will be a road closure in place on the Milton Interchange near Didcot with the entry and exit slip road closed from 9pm tonight until 6am.
The Milton Interchange work has been described by National Highways as “maintenance work”.
Motorists are being urged to plan their journeys in advance.
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It comes after the Hinksey Hill to Peartree route was also closed on Tuesday night which had an impact on those travelling to the Kassam Stadium as Oxford United beat Hull City 1-0 in a Championship fixture.
The works overran into Wednesday morning which had an impact on rush hour traffic.
National Highways said in a statement at 6.50am: “The A34 in Oxfordshire remains closed northbound between the A423 near Hinksey Hill and the Peartree Roundabout due to planned overnight roadworks that have overrun their pre-agreed end time due to issues with the tarmac.”
The road had reopened by 7.45am as National Highways thanked motorists for their patience as delays began to build northbound.