2009 Identification of the remains found in July 2007

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Proceedings of the National Academy of Sciences.
National Academy of Sciences

Genomic identification in the historical case of the Nicholas II royal family

  • Evgeny I. Rogaev,
  • Anastasia P. Grigorenko,
  • Yuri K. Moliaka,
  • Gulnaz Faskhutdinova,
  • Andrey Goltsov,
  • Arlene Lahti,
  • Curtis Hildebrandt,
  • Ellen L. W. Kittler,
  • and Irina Morozova
PNAS published online before print February 27, 2009, doi:10.1073/pnas.0811190106

Accurate unambiguous identification of ancient or historical specimens can potentially be achieved by DNA analysis. The controversy surrounding the fate of the last Russian Emperor, Nicholas II, and his family has persisted, in part, because the bodies of 2 children, Prince Alexei and 1 of his sisters, have not been found. A grave discovered in 1991 contained remains putatively identified as those of the Russian Royal family. However, not all family members were represented.

Here, we report the results of genomic analyses of new specimens, the human remains of 2 burned skeletons exhumed from a grave discovered in July 2007, and the results of a comprehensive genomic analysis of remains from the 1991 discovery. Additionally, ≈117 years old archival blood specimens from Nicholas II were obtained and genotyped, which provided critical material for the specific determination of individual identities and kinship identifications.

Results of genotypic analyses of damaged historical specimens were evaluated alongside samples from descendants of both paternal and maternal lineages of the European Royal families, and the results conclusively demonstrate that the recently found remains belong to children of Nicholas II: Prince Alexei and his sister. The results of our studies provide unequivocal evidence that the remains of Nicholas II and his entire family, including all 5 children, have been identified.

We demonstrate that convergent analysis of complete mitochondrial genome sequences combined with nuclear DNA profiles is an efficient and conclusive method for individual and kinship identification of specimens obtained from old historic relics.
Here is the press release from Dr. Coble, I post it with his permission:

US, European Scientists Report Findings of the Two Missing Romanov Children

An international team of researchers confirm the identity of the Tsar’s two missing children using forensic DNA testing.

Forensic DNA testing of skeletal remains of two individuals discovered in a field outside of Yekaterinburg, Russia in 2007 belong to Crown Prince Alexei Romanov and one of his sisters, ending one of the greatest mysteries of the twentieth century. The study, is published March 11th in the online, open-access, peer-reviewed journal PLoS ONE.

On July 17, 1918 Tsar Nicholas II, his wife Alexandra, their five children, the family physician, and three loyal servants were murdered by their Bolshevik executioners to prevent an attempted rescue from the nearby White Russian Army, who were loyal to the Tsar. After a botched attempt to hide the remains in an abandoned mine shaft, the Bolsheviks first tried to cremate two of the children (those discovered in 2007) and then buried the remaining nine bodies in a mass grave (officially discovered in 1991).

Dr. Michael Coble from the Armed Forces DNA Identification Laboratory (AFDIL) in Rockville, Maryland, was invited by the Prosecutor’s Office of Russian Federation to conduct an independent investigation of the remains in October of 2007. Coble and Dr. Anthony Falsetti, of the University of Florida, traveled to Yekaterinburg in November of 2007 to examine the remains. “Of the 44 bone fragments and teeth present, it was possible to determine that at least two individuals were present – one female and one putative male,” said Falsetti, a co-author of the study.

Coble and Dr. Odile Loreille, lead authors of the publication, also requested that another independent laboratory be involved. Dr. Walther Parson at the Institute of Legal Medicine in Innsbruck, Austria conducted an independent analysis of the remains in the spring of 2008. “It is important in an investigation of this magnitude that independent, parallel studies are conducted to confirm the results” according to Dr. Parson. In addition to the remains from 2007, both laboratories were given skeletal material from the 1991 for comparison. Both laboratories then blindly sent their results to Dr. Peter Gill at the University of Strathclyde in Glasgow, Scotland, for an independent evaluation of the results. Dr. Gill led the first DNA investigation of the 1991 mass grave in 1993.

Scientists from both laboratories used a three-prong approach to confirm their findings. First, mitochondrial DNA testing was conducted to confirm the sequence of the 2007 remains to the sequence of Alexandra and a living maternal relative, HRH Prince Philip, Duke of Edinburgh. Next, using sensitive techniques to amplify low levels of DNA, the scientists were able to develop a full Y-chromosomal STR profile from one of the bone samples that had been identified as a putative male. When compared to the Y-STR results from a tooth of Tsar Nicholas II, the researchers found an exact match. A comparison to a living Romanov cousin yielded the same Y-STR profile. Finally, the researchers developed a complete 16-marker autosomal STR profile for each member of the Russian royal family from the 1991 grave and from the two individuals in the 2007 grave.

The results demonstrate the presence of a family group, and represent the most comprehensive examination of the Romanov remains to date. “Here we are able to give a full account of all of the Romanov family and can conclude that none of the family survived the execution in the early morning hours of July 17, 1918,” the scientists note in the paper.

In addition to the analysis of the skeletal remains, Coble also collaborated with the Sverdlovsk Regional Forensic Bureau Laboratory in Yekaterinburg to confirm their independent autosomal STR and Y-STR analysis of a blood stain on a shirt worn by Nicholas when he was a young man. On April 29, 1891 while touring the city of Otsu, Japan the Tsarevich Nicholas Romanov was attacked by a Japanese policeman during an attempted assassination. Nicholas survived the attack and the bloody shirt he had worn that day was returned to Russia as a relic of the attack. The results were completely concordant. “For the first time, there is now a link between the ante-mortem evidence DNA profile from Nicholas II to the post-mortem skeletal remains from the first grave discovered in 1991,” said Coble.

“It’s rewarding to finally conclude this mystery 16 years after the first analysis of the remains,” reported Dr. Gill.

PLEASE ADD THIS LINK TO THE PUBLISHED ARTICLE IN ONLINE VERSIONS OF YOUR REPORT: http://dx.plos.org/10.1371/journal.pone.0004838 (link will go live tommorrow)

CITATION: Coble MD, Loreille OM, Wadhams MJ, Edson SM, Maynard K, Meyer CE, Niederstätter H, Berger C, Berger B, Falsetti AB, Gill P, Parson W, Finelli LN (2009) Mystery Solved: The Identification of the Two Missing Romanov Children Using DNA Analysis. PLoS ONE
DNA proves Bolsheviks killed all of Russian czar's children - CNN.com
CNN finally listing the same

I'll bet there are still some out there insisting otherwise. I remember reading a book that claimed all 5 children survived and had photos of family reunions through the years. I wish I could find a copy of that book again just to see what all it claimed. It is sad to get this confirmation of what most of us have believed all along--no survivors.
Thanks to Anna was Franziska, TRF members were able to read the report before it hit the news agencies. :flowers:
Yes! Thank you Anna was Franziska and TRF

Thanks to Anna was Franziska, TRF members were able to read the report before it hit the news agencies. :flowers:
A fine example of why I LOVE this site--facts, not only rumors and well versed opinions often backed with facts. Anna, I love your posts on other threads as well.
May the Romanovs now finally rest in peace.
That's good. I've hope that the family will be together soon.
I've moved discussion of the burial to a new thread in the main Russian Imperial Family forum.
Fragments of the pots that contained acid used on the Imperial Family have been found

Archaeologists are apparently still exploring the area around the old Koptyakovsky road (where the bodies were found) in Yekaterinburg for more clues as to what happened in July 1918. Yesterday, their efforts paid off when they found acid jags which contained the acid used in an attempt to dispose of the bodies and, in case the bodies were ever found, to make the verification impossible (as by the technology of the time).
The scientists believe that there can be no mistake as to the purpose of the acid in the vessels as the fragments were found in the same area as the bodies of the family. They also bear Japanese seals, as was typical for the acid jags sold in those times. So far, fragments of no less than 3 jags have been uncovered, each approximately 3-4 feet tall.
This complies with the accepted story of what happened to Nicholas II and his family in July of 1918.
The archaeologists continue searching the area in attempt to trace the actions of the assassins of the Imperial Family and cover any ‘white spots’ left.

Some fragments were found with the remnants of Alexei and Maria, however there were too few and small to carry out any analyses.

Russian-speaking members can read article about the new findings here.

It seems odd to me that they released the images of the original bones found, but few, if any, of the new bones. Also, since the bones were mere fragments, isn't it possible that these new remains were actually those of people who grew to adulthood and were planted in the forest after their deaths? Has anyone looked into possible age fraud of these new remains?
Also, since the bones were mere fragments
There's a reason why the bones are in small fragments, and that is because the bones have been in the grave for over nine decades.
Over much time, the bones deteriorated from sulfuric acid and weather conditions.

isn't it possible that these new remains were actually those of people who grew to adulthood and were planted in the forest after their deaths?
The theory about the bones being planted is not true. There's no evidence to prove that. These bone fragments have been accurately identified through the DNA evidence. Michael Coble from the world's largest mitochondrial DNA testing laboratory, along with the rest of the scientists spent huge amounts of time identifying these remains according to the reports that the Red Guard unsuccessfully tried to burn the remains of two of the dead children before burying them. This information matched well with the condition of the bones.

Has anyone looked into possible age fraud of these new remains?
There's no lying about the age of these bones. The scientists, used DNA to determine how old the remains are. The bones have been under the same soil for over 9 decades.You should watch the Finding Anastasia show that I've posted in the "Romanov children" thread. The whole documentary is all about the scientists observing and solving everything about the 44 bone fragments discovered on July 2007.
Okay... So... What gives here? Why is Georgy, of all people, now calling for a re-examination of the Ekaterinburg remains? What could
possibly be his motive?

From Today's Interfax
24 February 2010, 16:18
House of Romanovs wants royal remains to be re-examined

Moscow, February 24, Interfax - The House of Romanovs has said it wants to re-examine what is thought to be the remains of the last Russian Tsar Nicholas II and his family found in Yekaterinburg.

"Unfortunately, there is too much confusion in this case and some serious errors were made when the Yekaterinburg remains were being identified," Prince Georgy Mikhailovich, the son of the Romanovs' House head Grand Duchess Maria Vladimirovna, told Interfax on Wednesday.

"Many of the facts uncovered suggest that the Commission was staging a political show for a date, rather than working to establish the truth," Romanov said.

The remains were buried in St. Petersburg in 1998 in haste and in defiance of the opinion of the Russian Orthodox Church and the House of Romanovs, he said.

"No satisfactory answers have been provided to the ten questions, formulated by independent scholars and referred to the Commission back in 1998 by the late Russian Orthodox Patriarch Alexy II.

Meanwhile, those were questions, which interested not only experts, but also people and millions of believers. The Church needed to console these people instead of leaving them with a feeling of having been deceived. The Commission did not care. There was a farce instead of an act of repentance and purification," he said.

The remains' authenticity must be further studied, Romanov said.

Please See:
Why is Georgy, of all people, now calling for a re-examination of the Ekaterinburg remains? What could possibly be his motive?
His motive is to publicly align himself with the Orthodox Church's view, just as his mother has done previously.
It's not clear whether Georgi is questioning the 1994 Gill report of the DNA analysis, the 2009 University of Massachusetts Medical School 'Genomic Identification' report, or both.
Sorry guys, I've removed some posts that have no relevance to the 2009 'Genomic Identification' report (the subject of this thread).
Discussion of scientific and medical errors can be done in Members' Corner.

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