The grave contained nine sets of remains. Through a combination of anthropological analysis and DNA testing, scientists identified five of the remains as members of the Romanov family — Nicholas II, Alexandra, and three of their daughters — and the remaining four as members of the household staff who had been killed at the same time.
The DNA identification relied on mitochondrial DNA, which is inherited exclusively through the maternal line and therefore allows comparison across many generations of maternal relatives. Tsarina Alexandra’s mitochondrial DNA profile — shared by her children — could be compared against a living individual who shared the same maternal line. Prince Philip, Duke of Edinburgh, was that individual: his maternal grandmother was Princess Victoria of Hesse, sister of Tsarina Alexandra. Prince Philip donated a blood sample for the comparison.
The results confirmed that the bones in the Ekaterinburg grave belonged to Nicholas II, Alexandra, and three of their five children. The identification was supported by the mtDNA comparison with Prince Philip and by comparisons involving other living relatives for the paternal line.
A significant complication emerged: only three daughters were represented in the 1991 grave. The remains of Alexei, the Tsarevich, and one daughter were not found there. Which daughter was missing was a matter of genuine scientific dispute: anthropological analysis had suggested Anastasia or Maria, and the two camps — Russian researchers and American researchers — reached different conclusions. Russian scientists concluded the missing daughter was Maria. The American team concluded it was Anastasia.
This uncertainty — which remained unresolved through the 1990s and into the 2000s — had direct implications for the Anderson case. If Anastasia’s remains were among the 1991 find, the claim was definitively dead on that ground alone. If they were not, the biological evidence from the grave did not rule out a survivor.