The newly structured competition will see the 12 Guinness Premiership clubs and four Welsh Regions play over four weekends, followed by the semi-finals and finals. Each team will be guaranteed two home and two away inter-pool fixtures, with the top teams from each pool qualifying for the knockout stages.
Terry Burwell, Rugby Football Union Tournaments and Competitions Director, said: "We are delighted that together with Premier Rugby Ltd and the Welsh Rugby Union we have come up with a structure that will not only help players, coaches and officials develop but also provides more revenue potential for the Guinness Premiership clubs and the Welsh regions.
"There is a massive appetite for Cup rugby in England and Wales. The average attendance at pool matches over the past three seasons has been 8506 and the final averaged 60,000. There is plenty to look forward to with pool matches such as Gloucester Rugby against Cardiff Blues and Leicester Tigers against Newport Gwent Dragons in the first two rounds this coming season.
"We believe this new structure will give players, coaches and supporters an exciting tournament that embodies all that's best about cross-border knockout competition.
"We have a committed broadcaster in SkySports for the next five years and, together with PRL and the WRU, are actively seeking a title sponsor."
Leicester Tigers, Sale Sharks, Saracens, Ospreys
London Irish, Gloucester Rugby, Newcastle Falcons, Scarlets
Harlequins, London Wasps, Worcester Warriors, Cardiff Blues
Bath Rugby, Northampton Saints, Leeds Carnegie, Dragons
* Teams in Pool 1 play teams in Pool 4 and teams in Pool 2 play teams in Pool 3.
* Each team guaranteed four pool matches - two home and two away - with the top teams from each pool qualifying for the semi finals.
* Teams have been seeded depending on their finish in last season's Guinness Premiership and Magners League and best efforts have been made to avoid fixture duplication with the Heineken Cup in 2009-10.
Round 1 (weekend of November 7, 2009)
Leeds Carnegie v Leicester Tigers
Newport Gwent Dragons v Sale Sharks
Saracens v Bath Rugby
Ospreys v Northampton Saints
Gloucester Rugby v Blues
London Irish v London Wasps
Worcester Warriors v Newcastle Falcons
Harlequins v Scarlets
Round 2 (weekend of November 14, 2009)
Leicester Tigers v Newport Gwent Dragons
Sale Sharks v Leeds Carnegie
Bath Rugby v Ospreys
Northampton Saints v Saracens
Cardiff Blues v London Irish
London Wasps v Gloucester Rugby
Newcastle Falcons v Harlequins
Scarlets v Worcester Warriors
Round 3 (weekend of January 30, 2010)
Leicester Tigers v Bath Rugby
Sale Sharks v Northampton Saints
Newport Gwent Dragons v Ospreys
Leeds Carnegie v Saracens
London Wasps v Scarlets
Cardiff Blues v Newcastle Falcons
Gloucester Rugby v Worcester Warriors
London Irish v Harlequins
Round 4 (weekend of February 6, 2010)
Northampton Saints v Leicester Tigers
Bath Rugby v Sale Sharks
Ospreys v Leeds Carnegie
Saracens v Newport Gwent Dragons
Scarlets v Cardiff Blues
Newcastle Falcons v London Wasps
Worcester Warriors v London Irish
Harlequins v Gloucester
Semi finals (weekend of March 13, 2010, venue TBC)
Final (weekend of March 20, 2010, venue TBC)
* If the winner of the competition is an English club, they will qualify for the following season's Heineken Cup.