By conducting DNA analysis on the bones and the known living relatives of King Richard III, researchers have found compelling evidence that the skeleton unearthed by University of Leicester archeologists from a car park in 2012 indeed belonged to the last king of the House of York concluding history's oldest forensic investigation.

Dr. Turi King, from the University of Leicester Department of Genetics, and colleagues collected DNA from the living relatives of the 15th century king and conducted an analysis on several genetic markers including the mitochondrial genomes and Y-chromosomal markers from both the skeleton and Richard III's living relatives.

The mitochondrial DNA is inherited from the mother. Thus, the researchers were able make a comparison of the results of the DNA analysis of the skeletal remains and two living relatives of Richard III by following the female line of the king's family tree.

The Y chromosomes, on the other hand, are passed on by the father so the monarch would have inherited his father's, King Edward V, as would his present day relatives.

The researchers found a match between the mitochondrial DNA of Richard III and those of his living female-line relatives, Michael Ibsen and Wendy Duldig, supporting claims that the skeletal remains indeed belong to England's last king who died in a battle.

"Here we report DNA analysis of both the skeletal remains and living relatives of Richard III. We find a perfect mitochondrial DNA match between the sequence obtained from the remains and one living relative, and a single-base substitution when compared with a second relative," King and colleagues wrote in their study, which was published in the journal Nature Communications on Dec.2.

The researchers, however, did not find a match when they tried to trace Richard III's lineage of the Y chromosome. Richard III did not have the same Y chromosomes as his five living relatives, suggesting that infidelity may have occurred along the royal bloodline. It also raises questions about the legitimacy of Richard III's claim to the throne and those of later kings.

Nonetheless, the DNA analysis proved it was Richard III's skeleton that was buried and dug by archeologists from a parking lot in in Leicester two years ago. The researchers said that even with conservative estimate, the skeletons are confirmed to belong to Richard III with 99.999% accuracy.

"Even with our highly conservative analysis, the evidence is overwhelming that these are indeed the remains of King Richard III, thereby closing an over 500 year old missing person's case," King said.

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