"Given the fact that the necessary changes we seek to the backstop have not been secured between the Government and the European Union, and the remaining and ongoing strategic risk that Northern Ireland would be trapped in backstop arrangements at the end of the implementation period, we will not be supporting the Government if they table a fresh meaningful vote," the DUP said in statement late on Wednesday.
The party noted that it had discussed the need to maintain the United Kingdom's economic and constitutional integrity after Brexit with the government.
The DUP suggested that the backstop arrangement, agreed upon by London and Brussels, might lead to a hard border established on the island of Ireland, cutting Northern Ireland from its main market, which is the United Kingdom.
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The DUP's statement has provoked speculations on whether the party would abstain from the vote on the divorce deal.
Nigel Dodds, the DUP deputy leader, wrote on Twitter on Wednesday that the party members "do not abstain on the Union."
The statement comes shortly after May promised to resign if her fellow Conservative lawmakers back the withdrawal deal.
The no-deal scenario was defeated in a 160-400 vote in an indicative vote late on Wednesday.
The deal was agreed by the European Union and the United Kingdom last year and the United Kingdom was supposed to leave the bloc as soon as on March 29. However, UK lawmakers have failed to adopt the agreement, which sparked speculations about the possible no-deal withdrawal.
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Last week, the European Council approved the Brexit deadline extension, requested by UK Prime Minister Theresa May. Under the arrangement, the withdrawal would be delayed until May 22, if the divorce deal is approved by lawmakers by the end of this week; or until April 12, if the deal is not passed this week.