Solving twentieth century’s greatest mystery using DNA profiling — The case of the missing Romanov family members
By Niharika Adak
During the fall of 1917, Russia’s provisional government was overthrown by the Bolsheviks, which marked the beginning of the Bolshevik revolution. Following this the last ruling Russian Tsar, Nicholas II, abdicated his throne. On the night of July 16–17, 1918, Tsar Nicholas II and his wife, the Tsarina Alexandra, their five children (four daughters and one son who was the crown prince) and four loyal members of their staff were murdered by the Bolsheviks in Yekaterinburg, Russia.
The massacre in question was a big political move and hence was carefully planned. The Bolsheviks had been preparing for this day by loading the house (murder site) with benzene to burn the corpses, and sulfuric acid to disfigure them beyond recognition. To cover up the murders, the bodies were taken to the forest; covered in acid and buried. They killed each and every member of the Romanov family and dumped them in graves.
These murders gained prominence in 1991 when the graves were discovered accidentally. Nine skeletons were found in the grave — six adults and three children — The bodies showed signs of violence, consistent with reports of their treatment during and after death. Some of the remains were clearly aristocratic as their teeth were filled with porcelain, silver, and gold, dentistry well beyond the means of the average Russian, in the early 20th century. What most people wanted to know was if all the bodies belonged to members of the Royal Romanov family alone?
To answer this question, DNA was extracted from the bones. DNA is a stable genetic material that can be recovered from dead bodies long after the body has decomposed. Combining this DNA with specific reagents yields what are called short tandem repeats (STR). STRs are important DNA markers. This is comparable to the occurrence of short stretches as toll gates on a highway. These spots are indicative of which region one is looking at on the DNA just like how the toll gate indicates location within a state/city. Using these short stretches of DNA, one can identify similarities between parents and their children, siblings and relatives. Since the children’s DNA is a composite of the parents, a similar band pattern will be observed in specific regions. To confirm that the bodies belonged to the Royal Romanov family, DNA from the living members of the lineage were used to cross-verify the claims.
In this specific case, the DNA obtained from living relatives was the mitochondrial DNA. Mitochondrial DNA is contained in the energy-generating mitochondria of cells; it has the unique characteristic of being inherited solely from the mother and is transmitted maternally across generations.
The living relative whose mtDNA was collected for comparison was Prince Philip who belonged to the same lineage. On comparing, it was found that the mitochondrial DNA sequences from four of the female skeletons — the three children and the adult female identified as the Tsarina were an exact match implying that the bodies were all of the royal Romanovs.
However, only the bodies of Tsar Nicholas II, his wife, their three daughters and their four servants were identified. The bodies of the other two children- the only son and one daughter were still missing. In the anticipation of finding these bodies, the Sverdlovsk Regional Forensic Bureau Laboratory (Yekaterinburg) expected the possibility of future DNA testing, and preserved a limited number of fragments from each skeleton.
In the summer of 2007, a second grave 70m away from the first, was discovered by archaeologists. This grave contained a set of 44 bone fragments and teeth which were suspected to be that of the missing children. To confirm this, MtDNA matching was performed again. The DNA profiles of the two samples from the second grave fit perfectly into the family tree of the Tsar and Tsarina proving that they were indeed the missing children.
For close to about ninety years the disappearance of the Romanov family was shrouded in mystery. It wasn’t until several years after their execution that the Soviet government finally acknowledged the death of all of the Romanovs, thanks to the DNA fingerprinting technology.
The fact that a miniscule percentage of variability is what contributes to the uniqueness of a family tree is simply mind blowing! The same when put into practice has revolutionized the science of Forensics to a large extent. The scientific understanding of the structure and function of DNA and its stable inheritance, has helped unravel many mysteries. Starting from bone analysis for identification to using dried blood stains, hair and other body fluids to catch the killer, science has helped forensics come a long way! Cases like that of the Romanovs helped the world understand the importance of forensic evidence and the beauty of DNA that is present within each one of us.
About the Author — Niharika Adak is currently pursuing B.Tech in Biotechnology at VIT Vellore. She is an enthusiastic learner with a keen interest in cancer research. She is intrigued by the beauty of science and how it provides a solution for every problem. Connect with her on LinkedIn: Niharika Adak, Instagram: @niharika._.adak and Twitter: @AdakNiharika.
