Over 70 years after the night Tsar Nicholas II and his family were killed, the investigation into their murders was still open. No-one had found their bodies. But in 1991 some remains were excavated close to where they died, and a new part of the investigation began.
At this time I was working at the UK’s Forensic Science Service, the premier laboratory working on forensic identification with DNA technology. In 1984 Alec Jeffreys provided the first demonstration of DNA profiling, which showed people have DNA characteristics unique to them, just like fingerprints. A year later, in 1985, I worked with Alec at Leicester University on the first demonstration of DNA profiling applied to forensic-type materials like old blood stains. We also demonstrated that we could analyse very small or degraded samples of DNA to find someone’s DNA fingerprint.
I first met Russian scientist Pavel Ivanov at a scientific conference in 1992, and he asked me if I was interested in working with him to study the remains using DNA profiling techniques. I was interested, but this kind of study had never been done before and we didn’t know if it was possible to achieve any results from such old samples. Bones that had been buried for over 70 years would be very difficult to extract DNA from, as it degrades over time. But I knew this was an opportunity I didn’t want to turn down, and the UK Home Office supported our work.
I still remember meeting Pavel at Heathrow Airport arrivals – there was intense media interest and swarms of journalists were present. Pavel carried the bones from Russia and we transferred them to the Forensic Science Service, where the work began.
A small team of scientists worked with me and Pavel on the remains – Kevin Sullivan, Gillian Tully, Colin Kimpton, Nicola Benson and Romelle Piercy. We thought that the main problem we would face was ‘contamination’. Since the bones were discovered they had been handled by lots of different people. DNA is present in sweat, skin cells and saliva, so their DNA could be present on the bones as well.
We developed strict quality protocols to deal with this. We all wore clean lab coats and facemasks to prevent saliva spray while talking. To remove any modern DNA from the surface of the bones we filed them with sandpaper and took a small section of bone for analysis. As expected, we could only find very small amounts of DNA present in the bones. Each sample contained DNA originating from just half a dozen cells.
To determine if the remains belonged to the Romanovs, we needed to compare them to samples from verified relatives. We were fortunate to obtain blood samples from HRH Prince Philip, Duke of Edinburgh, who is a direct descendant of the Tsarina Alexandra. Samples were also obtained from the Duke of Fife and Princess Xenia Cheremeteff Sfiri, who are related to the Tsar. The remains matched their living royal relatives and we therefore knew we had found the bones of the Romanovs.
But this wasn’t the end of the story. Other scientists were surprised we’d been able to obtain any DNA from such old remains, and a very small section of the Tsar’s DNA sequence didn’t match his living relatives. We worked for another year to verify our results, but some still considered our findings controversial. A number of different groups of scientists in the USA and Russia worked to confirm or discount our results. One group even exhumed the body of the Tsar’s brother, George, from the St Petersburg cathedral. But each new test confirmed our original findings.
Our work identifying the Tsar’s remains helped to create the UK national DNA database and accelerated the development of new methods for forensic testing with small samples of DNA. Today these are used around the world in forensic investigations by the police and have been used to solve thousands of criminal cases.
This blog was written alongside our free exhibition The Last Tsar: Blood and Revolution, exploring the life and death of Tsar Nicholas II and his family and the forensic investigation into their murder. Discover more about the science behind one of the greatest mysteries of the 20th century in this series of blog posts.