Arguments have raged over whether some of the family survived the Bolshevik firing squad and conspiracy theories, some involving the British royal family and the Vatican, have abounded.
Even the bones found in Ekaterinberg two years ago were dismissed as a hoax by a conference of the Russian Union of Scientists in St Petersburg last month.
The DNA analysis by the Home Office Forensic Science Service has destroyed that criticism. The bones said to be those of the Tsarina and the three grand duchesses are related to the Duke of Edinburgh while those alleged to be of the Tsar are related to two of his living descendants.
Yet despite the scientific near certainties expounded by Dr Peter Gill and his Russian colleague Dr Pavel Ivanov yesterday, the most intriguing mystery of all, that of Grand Duchess Anastasia, remains unsolved.
In 1920 Anna Anderson surfaced in Berlin claiming to be Anastasia and her accurate recollections of life with the Russian imperial family convinced many people that she was genuine. She died in America in 1984 and last year a lock of her hair was sent to Home Office scientists who analysed the bones found in Ekaterinburg.
The hair has not yet been analysed. Dr Janet Thompson, director-general of the Forensic Science Service, said yesterday: 'We have not done any work on the pretenders as such. That possibility is under discussion.'
Analysis of the hair may prove or disprove Dr Anderson's claim. Blood tests could do the same for other claimed descendants such as the self- styled Prince Alexis II in Spain, who says he is the son of Maria, another of the four grand duchesses.
The Russian government must now decide whether to permit an imperial funeral for the bones in St Petersburg.
Prince Rostislav Romanov, grandson of the Tsar's sister, said yesterday: 'Our family is generally of the view that it would be better to bury them in Ekaterinburg where they were martyred and it should be made a monument to all the people murdered by the Communists.'