The choir will be seated in the north stand where they will be conducted by the stadium's feature choirmaster, Haydn James.
Some supporters' favourites such as Cwm Rhondda, Gwahoddiad , Calon Lan and Sospan Fach will feature in the repertoire.
The choir supported by the band of the Royal Regiment of Wales will be in place and on song from the moment fans start pouring into the stadium ahead of the 2.30pm kick off.
The 1,000 voices will also lead the singing during the match to make sure the stadium rings out to the traditional sound of Welsh rugby as the game is played.
Haydn James said:"This should give a spectacular sound from some of the very best choirs in Wales.
"We are all absolutely thrilled that so many choirs will be sharing in this very special day."
The full list of choirs is:-
Llanelli Male Voice Choir, Dunvant Male Choir, Morriston Orpheus Choir, Cardiff Arms Park Male Choir, Cor Meibion Taf, Pendyrus Male Choir, Bechgyn Bro Taf, Treorchy Male Choir, Cor Meibion De Cymru, Froncysyllte Male Choir, Cor Dodre'r Aran, Cor Meibion Llangwm, Cantorion Colin Jones, Built Wells Male Choir, Cor Meibion Maesteg, Maesteg Gleemen, Cor Yr Hen Blwyf, Glynneath Male Choir, Cor y Gyrlais, Eschoir, Cor Meibion Dyffryn Aman and Risca Male Choir, Cambrian Male Choir and Abercynon Male Voice Choir.
The Millennium Stadium Venue Sales Manager, Alex Luff, said:"This has to rank as one of the biggest choirs ever recruited to support a major rugby international.
"The sound will be superb and will undoubtedly add to the emotion and atmosphere of a great occasion.
"We are renowned as a land of song and it is only fitting that we honour that heritage on the day the Welsh Rugby Union celebrates its 130th anniversary."