Since one of the finalists, Shakhtar Donetsk, has already qualified for European competitions for 2009–10 by virtue of their position in the Premier League, Vorskla Poltava automatically qualifies for the 2009–10 UEFA Europa League.

All draws held at FFU headquarters (Building of Football) in Kyiv unless stated otherwise.

In this round entered 25 teams from Druha Liha and winners of the Ukrainian Amateur Cup.

They were drawn against the 14 winners of the First Preliminary Round, with two matches containing only Persha Liha teams.

Notes: The Ukrainian Cup Final was played in Dnipro Stadium, Dnipropetrovsk on May 31, 2009.