BRITISH forensic scientists jubilantly announced yesterday that they have identified bones found in Russia two years ago as those of the last tsar and tsarina and three of their daughters.
They are almost 99 per cent certain that they have solved the mystery of the fate of the Russian imperial family. But enough questions remain unanswered to keep the arguments and conspiracy theories going for years.
One of the daughters, possibly the Grand Duchess Anastasia, and the Tsar's heir, Alexei, were not among the skeletons found in a pit at Ekaterinberg in the Urals, where the family was murdered by the Bolsheviks 75 years ago next week.
Nevertheless, yesterday's announcement was both historically important and a triumph for the DNA testing techniques pioneered by British scientists. Dr Janet Thompson, director-general of the Home Office Forensic Science Service, called it 'a proud and exciting moment'.
After nine skeletons were discovered in the pit in 1991 the Russians, using anthropological and dental tests, tentatively identified them as Tsar Nicholas II, Tsarina Alexandra, three children, the royal doctor and three servants.
The Forensic Science Service was approached by the Russians and asked to provide more definitive identifications. Bone samples were brought to Britain last September and their cells analysed for deoxyribonucleic acid (DNA), which makes up a person's unique genetic blueprint.
Dr Peter Gill, who led the research team, said yesterday: 'We are more than 98.5 per cent certain that the remains are those of the Romanovs.'
Dr Gill and his colleagues took blood and hair samples from the Duke of Edinburgh and two unnamed descendants of the Tsar. The Duke's maternal grandmother was Princess Victoria of Hesse, the Tsarina's sister.
The bones were tested for mitochondrial DNA, passed down through the maternal line and present in larger quantities than chromosomal DNA. A precise match between the Duke's DNA and that taken from the remains of the Tsarina and the three children was obtained.
But the DNA in what was thought to be the Tsar's bones did not quite match that of his two descendants, even though the disparity was only one part in 800. The research team at Aldermaston, Berkshire, was not satisfied.
They therefore amplified the Tsar's DNA using a process known as polymerase chain reaction. Because there is then more DNA the tests are more sensitive and the research team solved the problem.
Dr Gill said: 'We found what we suspected, that there were two different types of molecule in the clone.
'One was exactly the same as the descendants and the other was the same as the apparent anomaly. Type two was the result of a rare mutation.'
To establish that the Tsar was the father of the three children in the grave the scientists analysed chromosomal DNA for what is known as short tandem repeats. This showed that the three girls had inherited DNA from the Tsar.
Further evidence such as the gold and platinum dental fillings that only the rich could afford, wounds on six skeletons, bullets from a revolver of the right age, and the fact that the Tsar's skeleton was the correct height, convinced the team these were the Romanovs.
But the Tsar and Tsarina had four daughters and the scientists were not able to tell which one was missing from the grave because the girls shared similar DNA codes.
The mystery of whether Anastasia escaped the massacre remains unsolved and the truth of the claims of the late Anna Anderson to be the missing Grand Duchess is unresolved. A lock of her hair has not yet been analysed.
The nine skeletons definitely did not include the Tsarevich Alexei, the heir to the throne. According to some historical accounts his body and that of Anastasia were taken away and burnt.
Grand Duchess mystery, page 3
Join our commenting forum
Join thought-provoking conversations, follow other Independent readers and see their replies