DNA Profiling Module Writing Assignment

DNA Profiling Part 2

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Part 2 of DNA Profiling: the case of the Romanovs

•  The last Tsar of Russia, Nicholas II had a wife, Alexandra, four daughters, Olga, Tatiana, Maria and Anastasia and a son, Alexei

•  After a lengthy period of civil unrest, and amidst the devastation of WWI, Nicholas II abdicated his throne in March, 1917

•  First the provisional (Karensky) government was in power for a few months but in October 1917 the Bolsheviks took over during the Bolshevik Revolution

Exile and Murder •  The ex-Tsar and his family

were eventually imprisoned in the “Ipatiev House” in Ekaterinburg, a small town in the Ural mountains

•  On July 17, 1918 the family was taken to a basement room of the house and murdered

•  The locations of the bodies were a Russian state secret for about 70 years

•  There were persistent rumors throughout the 20th century that two of the children, Alexei and Anastasia, had possibly survived.

Ipatiev House

Room where murders took place

romanov-memorial.com

awesomestories.com

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Did one of the daughters survive?

•  Anna Anderson (Manahan) appeared mysteriously in 1921 and claimed to be the youngest daughter of the Tsar, Grand Duchess Anastasia

•  She looked a lot like Anastasia, was about the same age, and spoke fluent Russian and German

•  She also knew details of Anastasia’s life that were not widely known outside the Romanov family circle

•  Some believed her to be Anastasia even to her death in 1984 including Tatiana Botkin the daughter of the Romanov family doctor.

Gravesites found near Ekaterinburg were determined to be those of the Tsar and

his family The main grave found in 1991 contained remains of Tsar, Tsarina, 3 daughters and 4 servants

Another grave found in 2007 (a few hundred meters away) contained remains of the son and one of the daughters http://www.history.com/topics/russian-revolution/videos/ finding-the-romanovs

romanov-memorial.com awesomestories.com

Dr. Alexander Avdonin: discoverer of 1st grave

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Forensic work included analysis of bone fragments, dental records, blood samples and DNA

romanov-memorial.com

*Rogaev, et al., PNAS, 2009, 106(13), pp. 5258-5263.

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* awesomestories.com

Mitochondrial DNA sequencing helped determine the identities of the skeletons

found in the second grave •  Mitochondrial DNA is

passed only from mother to child

•  There must be a chain of “unbroken maternal lineage” established to use mtDNA to link two people genetically

•  But a son’s mtDNA can be compared to his mother, grandmother, great grandmother, etc.

•  The red or black filled circles and squares represent familial relationship groups that can be confirmed by mitochondrial DNA sequencing

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•  DNA data show that skeletons from both graves match each other as well as maternally-linked descendants from Queen Victoria

Genetic Marker Database Alexei Maria Tsarina Descendants

Across a row the sequences are identical.

•  Y-STR DNA analysis allows comparison of paternally-linked family members

•  Y-STR analysis showed that the skeleton believed to be the son, Alexei, in one grave matched the skeleton believed to be the Tsar found in the other grave as well as DNA from a bloodstained shirt.

Tsar Alexei

Paternal DNA Testing

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*They still had a shirt worn by Nicholas II during an assassination attempt in Japan in 1891 that they knew had his blood on it! The stain on the shirt was 117 years old!!!

The “allele” is the same for all samples at this locus, consistent with

them deriving from paternally related

individuals.

Many more alleles coming up the same

strengthens the conclusion the two

individuals were related.

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Okay, so what about the identity of Anna Anderson?

•  Anna Anderson (Manahan) appeared mysteriously in 1921 and claimed to be the youngest daughter of the Tsar, Grand Duchess Anastasia

•  Tissue and hair samples from the deceased Anna Anderson Manahan were compared to living relatives of the Romanovs and DNA from the Tsar and Tsarina’s skeletons found near Ekaterinberg

•  The samples were also compared to a living relative of Franzisca Schanzkowska, a polish ammunitions factory worker who disappeared about the same time as Anderson appeared in Berlin

DNA analysis of tissue samples from Anna Anderson as compared to Tsar and Tsarina

•  Are results from any one of the six loci consistent with her BEING the Grand Duchess Anastasia?

•  Are results from any of the loci consistent with her being a child of EITHER the Tsar OR the Tsarina? Which ones? Why?

•  Are results from any of the loci inconsistent with her being the child of either the Tsar or Tsarina? Which? Why?

Locus 1 2 3 4 5 6

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DNA analysis of tissue samples from Anna Anderson as compared to a relative of the Tsarina and a relative

of Franzisca Schanzkowska, a WWI munitions worker from Poland

•  Does this data establish her as directly related to Carl Maucher (great nephew of Schanzkowska)?

•  Does this data rule out that she is related to the Tsarina?

•  Does it matter that the “blood lines” are all maternal?