Of the children of Nicholas II escaped no one

titleRussian and American genetics, conducted a DNA analysis of human bones discovered near Yekaterinburg and tentatively identified as the remains of the family of the last Russian Emperor Nicholas II.The results of the analysis clearly showed that the remains actually belong to Nicholas, his wife Alexandra and their five children. Thus, finally it is proved that the entire Royal family was murdered in 1918, and none of them survived.An international group of scientists under the leadership of Eugene Rogaev from the University of Massachusetts and the Institute of General genetics, Russian Academy of Sciences reported in the journal Proceedings of the National Academy of Sciences of the USA on the results of the genetic analysis of the remains of the family of Nicholas II. Readers of "Elements" are already familiar with the works Rogaev and his colleagues note, "a Fully decoded mitochondrial genome of the mammoth" ("Elements", 07.02.2006). Worked on the mammoth methods of extraction and analysis of DNA fragments from very poorly preserved specimens now useful to scientists to address important historical questions.In the grave, was found in 1991 near Yekaterinburg, were found the remains, identified by forensic examination as belonging to Nicholas II, his wife Empress Alexandra, their daughters Olga, Maria, Tatiana, and three servants, and court physician. The remains of two children, Alexei and Anastasia - were not found, which became the basis for different assumptions and speculations (however, rumors that Alexei and Anastasia could be saved, have been circulating a long time). In July 2007, was the second grave was found, which drew 44 bone fragments, badly burnt and damaged - apparently, sulfuric acid. Preliminary anthropological analysis showed that the bones belong to a boy 10-14 years old and the girl 18-23 years. In his article, Rogaev and his colleagues make a compelling genetic evidence for the affiliation of these remains of Alexei and Anastasia.Scientists were able to isolate DNA from two relatively well-preserved bone fragments from the second grave. Were included in the analysis also the bone fragments purportedly belonging to Nicholas, Alexandra and their three daughters from the first grave. For comparison used was the genetic material of living relatives of the Royal family: the descendants of Queen Victoria (grandmother of Alexandra and Empress Maria Feodorovna (mother of Nicholas II) in the female line, and Nicholas I in the male line.First, the researchers identified two bone fragments of the so-called hypervariable sites of mitochondrial DNA, which is easiest to carry out genetic identification, and compared them with the corresponding sections of the mtDNA from the bones allegedly belonging to Alexandra and her three daughters. For greater reliability analyses were conducted independently in two laboratories. All six tsakonian hypervariable sites of mtDNA was identical. As you know, mtDNA is passed only through the maternal line, so this result testified in favor of the fact that in the second grave contained the remains of children of Alexandra.The next step was the reading of the complete mitochondrial genomes of two bone fragments from the second grave. Have also read the complete MT-genomes of Alexandra and two other descendants of Queen Victoria on the direct maternal line. All these MT-genomes (length 16 571 nucleotides) were identical.To find out how prevalent in human populations this variant of the MT genome, the researchers turned to genetic databases that contain sequences of hypervariable sites of mtDNA over 70,000 people. Among them there was only one person having the same hypervariable area, like Queen Victoria and her descendants in the female line. This man had found, was otdekorirovana other parts of the MT genome and found a few differences. Thus, the MT-genome characteristic of the descendants of Queen Victoria, proved to be unique for the studied sample.Comparison of mtDNA from the alleged remains of Nicholas II with other mtDNA descendants of Maria Feodorovna confirmed that the remains actually belong to her direct descendant through the maternal line. Maria Feodorovna had heteroplasmic mutation at position 16169 of the mitochondrial genome. In other words, she had two types of mitochondria: cytosine and thymine at this position (16169C/T). Nicholas has inherited both types of mitochondria, whereas in the investigated descendants of his sisters were only one of two types (either C or T).The researchers were able to extract from the bones and also some parts of the nuclear genome. This allowed to show that one of the two bone fragments from the second tomb belongs to a man and the second woman. For sex determination, we used two independent methods (amelogenin gene and loci on the X and Y chromosomes, located at a distance of 6-8 million nucleotide pairs from a gene). These same methods was confirmed and gender of all of the remains from the grave.There was also an analysis of the fragments of Y chromosome selected from the alleged remains of Nicholas and Alexei. They were, first, identical, second, completely aligned with the corresponding sections of the now living lineal descendants of Nicholas I in direct male line, thirdly, they were unique, that is not met anywhere else in the available genetic databases.Further analysis of nuclear DNA fragments confirmed that the remains of graves from the second:1) belong to a man and a woman;2) do not belong to any of those whose remains are found in the first grave;3) directly related through paternal and maternal lines with the individuals from the first grave, believed to belong to Nicholas and Alexandra.Scientists have also examined the traces of blood on the shirt of Nicholas II from the archives of the Hermitage. This shirt has been preserved as a historical relic after the assassination of Nicholas II, which occurred in 1891 in Japan. Fragments of mitochondrial and nuclear DNA extracted from the blood, completely coincided with those that were previously allocated from the bones of the Emperor, were found in the first grave. In particular, in the blood were the same two options of mitochondria with cytosine and thymine at position 16169. This suggests that heteroplasmy (the presence of mitochondria with different genomes) Nikolai was not tissue-specific phenomenon and was observed in bone tissue and in blood, and two types of mitochondria in both cases received in the same proportion. All the obtained genetic data suggests that the bones of the first tomb, considered to be the remains of Nicholas II, really belong to the last Russian Emperor. The probability of error is completely negligible (about one chance out of 1025).As for the bones from the second grave, they, no doubt, belong to Alexei and Anastasia. The reliability of this conclusion is somewhat lower than in the case of the remains of Nicholas, but it is still quite astronomical: the probability that a man and a woman, whose remains are found in the second grave, are not children of Nicholas and Alexandra, is about 10-8-10-9.Thus, to date, the remains of Nicholas II, his wife Alexandra and all their five children was found and identified. From the family of the last Russian Emperor escaped no one. Source: children of Nicholas II escaped no one.

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