Russia cut shipments on January 1 after the dispute over prices stalled the renewal of the gas agreement.
Under a EU-brokered deal, Moscow had resumed supplies on Tuesday only to shut them off again hours later.
Gazprom, the Russian national energy provider, claimed that Ukrainians were refusing to allow gas transit under conditions laid down by the deal.
Naftogaz, the Ukrainian state-run gas company, said they would not resume deliveries due to "impossible" transit conditions set by Gazprom.
Neave Barker, Al Jazeera's correspondent in Moscow, said: "What Russia was keen to push across ... was that this was no longer a bilateral issue between the Ukraine and Russia.
"That's because the entire continent was involved and therefore European partners, those countries hard hit in eastern Europe and representatives from the European Union, should get involved in trying to end this situation."
The EU had warned Russia and Ukraine that its relations with them would be reviewed if the dispute was not resolved this weekend.
Jose Manuel Barroso, the EU commission president, said he would advise European companies to take legal action against the two countries if gas transit was not resumed.
Ukraine, an ex-Soviet state with pro-Western leaders, has also angered the Kremlin in the past months because of its attempts to join Nato.
Russia is the world's biggest natural gas producer and provides about a quarter of the gas used in the European Union - about 40 per cent of the gas the bloc imports.
About 80 per cent of the imports pass via pipelines in Ukraine.