The end result was that these two bodies were buried in one grave and the rest of the bodies in another, a fact that had fueled speculation that two of the children had managed to elude the bullets and escape.
Based on DNA technology available in the 1990s, the first five bodies were positively identified as the Tsar, the Tsarina and three of their children.
The second set yielded 44 bone fragments and teeth which were subjected to three types of genetic testing: mitochondrial DNA, autosomal STR and Y-STR testing.
The mitochondrial tests (mitochondria are passed through the mother) confirmed that the bodies were children of the Tsarina. Confirmation was done with a living maternal relative, HRH Prince Philip, Duke of Edinburgh.
The autosomal STR test was basically a paternity test, Coble said, revealing that the newer remains are 4.3 trillion times more likely to be related to the Tsar and Tsarina than two random individuals. Skeletal remains from the three daughters found in the first grave also matched up.
"It's as strong as evidence you're going to find as far as nuclear DNA goes," Coble said.
The Y-STR testing, done only on the remains of Alexei, matched the STR profile of the Tsar and also a living relative, Prince Andrew Romanov, a cousin of the Tsar.
The STR and Y-STR findings were confirmed by testing blood on a shirt that Nicholas had worn on a trip to Japan in April 1891, when he was attacked with a saber. The shirt had been stored at the Her
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