Remains of George Washington's long-lost grandnephews are identified using DNA from 200-year-old bones found in unmarked graves

Unmarked graves belonging to the grandnephews of the first US president have been identified after 200 years using DNA.

The bones belonged to Samuel Walter Washington and George Steptoe Washington Jr., along with their mother, Lucy Payne Washington.

The graves were discovered during a 1999 excavation at the Harewood Family Cemetery in Charles Town, West Virginia that aimed to find the remains of Washington's younger brother, Samuel Washington.

Researchers at the Armed Forces DNA Identification Laboratory (AFDIL) used DNA from the fragmented bones and a living descendent, Samuel Walter Washington.

This marks a significant discovery for the researchers who said their findings could lead them to identify service members extending back to World War II.

Researchers identified the remains of three of George Washington's ancestors using new DNA sequencing methods 

George Steptoe Washington Jr. lived from 1806 to 1831 and was buried in the Harewood Family Cemetery in West Virginia

Samuel Walter Washington lived from November 16, 1734 to September 26, 1781 and was a planter, politician and military officer buried at the Harewood Family Cemetery in West Virginia

In the study, the researchers had set out to identify all the unmarked graves at Harewood Cemetery and believed the recovered remains belonged to George Washington's descendants, but conducted DNA tests to be sure.

The researchers believed the recovered remains belonged to Washington's descendants because the burial ground sat on the land of the Harewood Estate, which was built by Samuel Walter Washington in 1977.

The most common form of DNA profiling is short tandem repeat analysis - which repeats DNA sequences to verify family connections - but it is often difficult, if not impossible, to use this method on degraded remains.

People who were embalmed using post-war techniques - such as Samuel, Lucy, and George Steptoe - involved formaldehyde which can damage the DNA.

Instead, the researchers performed a series of DNA tests that compared the remains to a living descendant: S.W. Washington. 

The researchers analyzed the Y chromosomes to determine the paternal relationship and mitochondrial DNA sequencing to determine the maternal relationships to build a family tree.

The grandnephews were more closely related than previously thought because of a cross-cousin marriage

The Herewood estate in Charles Town, West Virginia

The family of three was buried at the Harewood Family Cemetery in West Virginia

Mitochondrial DNA sequencing detects the smallest forms of biological material that is passed down from a mother to her child.

They also used a new method that analyzed DNA data from 95,000 sets of genomes which houses all the genetic information of a person.

'This particular case gave us an opportunity to test methods for extended kinship prediction that we developed using a set of known, degraded DNA samples needing identity confirmation,' said senior author Charla Marshall, Deputy Director of DoD DNA Operations. 

'Our laboratory is currently validating these novel methods to be used in routine casework.' 

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Through these methods, the researchers not only confirmed that Lucy, Samuel and George Steptoe Jr. were Washington's descendants, but they were surprised to find that their family tree was closer than they expected. 

The familial relationships were one degree closer, but the researchers were able to confirm that it was due to cross-cousin marriages in the Washington family tree. 

'Our data confirmed the identities of the three sets of remains, and we furthermore resolved which male was the direct ancestor of S.W. Washington, the living descendant,' Marshall added.

Samuel was Washington's brother and a planter, politician and military officer who lived from 1734 to 1781.

He was the county lieutenant of Berkeley County, Virginia during the early revolution and resigned in 1777 at 43 years old.

Little is known about George Steptoe II who lived from 1806 to 1831.

He was the youngest son of a Virginia planter and militia officer who died of Tuberculosis at 36.

This is not the first time researchers have had to use new DNA methods after scientists analyzed the hair of a man who claimed to be the great-grandson of Sitting Bull - a legendary Native American Chief. 

In 2021, the scientists used autosomal DNA from a lock of Sitting bull's hair to match it to Ernie Lapointe and found it was a positive match.

Now, uncovering Washington's grandnephews through new DNA sequencing could mean that formerly unknown and unidentified remains can be identified.

It will allow the researchers to expand the reference points to third and fourth-degree living relatives 'in an effort to increase the number of DNA-assisted identifications, particularly those of past conflicts such as World War II, Korea, Cold War, and Southeast Asia/Vietnam,' Marshall said.

Dailymail.com has reached out to the authors for comment.