British forensic scientists announce that they have positively identified the remains of Russia’s last czar, Nicholas II; his wife, Czarina Alexandra; and three of their daughters. The scientists used mitochondria DNA fingerprinting to identify the bones, which had been excavated from a mass grave near Yekaterinburg in 1991.

On the night of July 16, 1918, three centuries of the Romanov dynasty came to an end when Bolshevik troops executed Nicholas and his family. The details of the execution and the location of their final resting place remained a Soviet secret for more than six decades. Lacking physical evidence, rumors spread through Europe in the aftermath of the Bolshevik Revolution, telling of a Romanov child, usually the youngest daughter, Anastasia, who had survived the carnage. In the 1920s, there were several claimants to the title of Grand Duchess Anastasia. The most convincing was Anna Anderson, who turned up in Berlin in 1922 claiming to be Anastasia. In 1968, Anderson emigrated to Charlottesville, Virginia, where she died in 1984.

In 1991, Russian amateur investigators, using a recently released government report on the Romanov execution, found what they thought to be the Romanov burial site. Russian authorities exhumed human remains. Scientists studied the skulls, claiming that Anastasia’s was among those found, but the Russian findings were not conclusive. To prove that the remains were indisputably those of the Romanovs, the Russians enlisted the aid of British DNA experts.

First, the scientists tested for gender and identified five females and four males among the remains. Next they tested to see how, if at all, these people were related. A father and mother were identified, along with three daughters. The four other remains were likely those of servants. The son Alexei and one daughter were missing.

To prove the identity of Alexandra and her children, the scientists took blood from Prince Philip, the consort of Queen Elizabeth II and the grand nephew of Alexandra. Because they all share a common maternal ancestor, they would all share mitochondria DNA, which is passed almost unchanged from mother to children. The comparison between the mtDNA in Philip’s blood and in the remains was positive, proving them to be the Romanovs. To prove the czar’s identity, who did not share this mtDNA, the remains of Grand Duke George, the brother of Nicholas, were exhumed. A comparison of their mtDNA proved their relationship.

The Crown Prince Alexei and one Romanov daughter were not accounted for, adding fuel to the persistent legend that Anastasia had survived execution. Was it possible that Anastasia had escaped and resurfaced as Anna Anderson?

In 1994, American and English scientists attempted to answer this question once and for all. Using a tissue sample of Anderson’s recovered from a Virginia hospital, the English team compared her mtDNA with that of the Romanovs. Simultaneously, an American team compared the mtDNA found in a strand of her hair. Both teams came to the same decisive conclusion: Anna Anderson was not a Romanov. In 1995, a Russian government commission studying the remains presented what it claimed was proof that one of the skeletons was in fact Anastasia’s, and that the missing Romanov daughter was, in fact, Maria.