I’m excited to share the paper, “Unearthing Who and Y at Harewood Cemetery and inference of George Washington’s Y-chromosomal haplotype” by Cavagnino et al. 2024, and published in iScience, on which I’m a co-author.
When Goran Runfeldt, Head of R&D at FamilyTreeDNA called me last year and asked if I wanted to work on something fun, I had no idea of the significance of the journey I was about to undertake. I was privileged to join the team working on the Washington family story, as told through DNA via excavated family burials.
I’ll tell you upfront that this project is very close to my heart in a very personal way.
Let’s talk about the science first, then I’ll share my exciting personal connection.
The Washington Project
By the time I joined this study, Courtney Cavagnino and the team at Armed Forces DNA Identification Laboratory, a division of the Armed Forces Medical Examiner System (AFMES-AFDIL), had already been hard at work sequencing burials from the Harewood Cemetery in West Virginia for some time.
The Harewood Cemetery is located on a plantation owned by the Washington family where two grandsons of President George Washington’s brother, and their mother, Lucy Payne, are buried in unmarked graves.
George Washington’s brother, Samuel Washington (1734-1781), had the home designed in 1770 and had moved there before his death in 1781 at the age of 46, from tuberculosis. George Washington (1732-1799) visited his brother there several times.
Samuel Washington’s son, George Steptoe Washington (1771-1809), eventually inherited the property and married Lucy Payne (1769-1846). With Lucy, he had sons Dr. Samuel Walter Washington (1797-1831) and George Steptoe Washington II (1806-1831).
Lucy Payne’s younger sister, Dolley, married James Madison, the future President, in the parlor at Harewood in 1794.
This graphic from the paper shows Samuel Walter Washington’s ancestors. Note that he is related to Augustine Washington and Mary Ball through three different paths.
The FamilyTreeDNA research team redrew the relationships in a more traditional genealogical view.
Image courtesy FamilyTreeDNA. Click to enlarge.
Complicating the analysis, and making it more interesting was the fact that present-day tester, Samuel Walter Washington (SWW) is descended from Augustine Washington, the patriarch of the colonial Washington Family, and his wife, Mary Ball, through three different paths.
The Burials
According to the 1882 last will and testament of Dr. Samuel Walter Washington’s wife, the graves at Harewood were relocated to the Zion Episcopal Churchyard in Charles Town, West Virginia, where gravestones were placed for the Washington males. Therefore, only fragments and small bones were left in the Harewood plantation graves.
The Harewood property still remains in the Washington family, so they had ready access to the cemetery location. The original excavation took place in May of 1999, after using ground-penetrating radar to identify the likely burial locations based on soil disturbances. The original goal was to locate the grave of Samuel Washington, George Washington’s younger brother.
As would be expected, bacteria had contaminated already degraded DNA. This precluded traditional as well as some forensic sequencing methods. DNA capture technology has improved significantly since 1999, so the AFMES-AFDIL team was using a combination of revolutionary technologies to process the remains.
A technique known as hybridization capture using bait panels was combined with NGS sequencing to attempt to obtain about 95,000 nuclear SNPs, similar to those used in traditional autosomal testing. Additionally, the capture was primed for mitochondrial and Y-DNA SNPs for haplogroup determination. Some Y STRs were captured as well. The paper, published today, provides more technical details for those who are interested.
Three Kinds of DNA
We were fortunate to be able to utilize three types of DNA in the analysis.
Each type of DNA, with its specific inheritance characteristics, was critically important for establishing relationships between the burials. The connection to SWW identified the male burials.
- Y-DNA is passed only from male to male and is not mixed with the DNA of the mother, making it uniquely qualified for male lineage matching.
- Mitochondrial DNA is passed only from women to both sexes of their offspring, not mixed with the DNA of the father, making mitochondrial DNA uniquely qualified for matrilineal lineage matching.
- Autosomal DNA is inherited from all ancestral lineages and is divided in each generation. Half is inherited from one’s mother and half from one’s father. Based on both random inheritance and recombination, people, on average, inherit half the amount of autosomal DNA of each ancestor that their parents did.
Y-DNA
Y-DNA is passed from father to son intact, meaning that it is not mixed with the DNA of the mother. Small mutations accrue over time, forming branches of the Y-DNA phylogenetic tree. Those branches have names assigned, called haplogroups. The higher up the tree, the more descendant branches have occurred over time. The further down the tree, the more unique and refined the haplogroup. Haplogroups are formed when two or more men have the same group of unique mutations.
Additionally, a second type of Y-DNA, STRs, or short tandem repeats, is also used for comparison. These mutate much more quickly than SNPs, single-nucleotide polymorphisms, used to determine haplogroups. Both types of Y-DNA are utilized together.
The bait panels were constructed to recover at least some information about the Y-DNA of the male individuals buried in the graves. For comparison purposes, Samuel Walter Washington, the living descendant, took the highly refined Big Y-700 test at FamilyTreeDNA which tests millions of locations on the Y chromosome – including all of the locations on the bait panels..
Some Y-DNA of the two male burials was recovered and reconstructed. The DNA results matched each other, as would be expected of brothers, and also the Y-DNA of SWW.
This provided a relatively high-level haplogroup designation, R-U152, which was formed about 4500 years ago.
A matching haplogroup at this level does not confirm a close family relationship, but it also doesn’t preclude it.
Fortunately, the Big Y-700 test of SWW was able to reveal significantly more information, including his refined haplogroup of R-FTE201 which was formed about 2000 years ago.
George Washington didn’t have any known children, so we can’t compare his Y-DNA or autosomal DNA directly to either the Harewood burials or SWW.
Barring an unknown paternity event, George Washington’s Y-DNA haplogroup would be the same as that of his brother’s grandsons and the same as present-day tester SWW.
Of course, it’s possible that small mutational differences would have occurred in the past three centuries, since Augustine Washington, the common ancestor of George Washington and SWW, lived, but if so, their haplogroups would be nearly identical.
The Washington family has graciously permitted the Washington lineage to be included in Discover, so if you are haplogroup R, please check to see if the presidential Washington family shows up in your Notable Discover connections in the next few days.
Mitochondrial DNA
Mitochondrial DNA is passed from mothers to all of their children without being admixed with the father’s mitochondrial DNA. Only females pass it on. Therefore, to obtain the mitochondrial DNA of any ancestor, one must descend from that female ancestor through all females. In the current generation, the tester can be a male.
Mitochondrial DNA has been the chosen methodology for the identification and repatriation of military remains for at least two decades. The reason is simple. Mitochondrial DNA is easier to retrieve since thousands of copies live in the cytoplasm of each cell. Only one copy of the 23 pairs of autosomes lives in the nucleus of a cell.
The mitochondria are comprised of 16,569 locations, while the autosomes contain 3 billion pairs, for a total of 6 billion locations across both the maternal and paternal chromosomes. As you can imagine, degraded autosomal DNA is broken into small pieces and mixed together. Think of a blender. Recovering that DNA and then piecing it back together is a massive undertaking.
Furthermore, with military repatriations, the mother or sibling or other relative who shares the mitochondrial DNA of the soldier contributes their mitochondrial DNA to the military for comparison against remains as they are recovered.
One of the ways that the graves of Dr. Samuel Walter Washington and his brother, George Steptoe Washington, were confirmed is that the mitochondrial DNA recovered from those burials matches the mitochondrial DNA of another burial, which was determined to be their mother, Lucy Payne.
While mitochondrial DNA alone is generally not adequate to definitively prove identity, it can be utilized along with other evidence, such as extra mutations in addition to haplogroup-defining mutations, and the geographical location where the remains were recovered.
The AFMES-AFDIL team recovered the full sequence of Lucy Payne’s and her sons’ mitochondrial DNA, which was identified as haplogroup J1c1b1a1 based on unique haplogroup-defining mutations.
Why the AFMES-AFDIL Team?
You may recall that the US government agency involved in this project is the Armed Forces DNA Identification Laboratory. Why, you might wonder, are they involved in the identification of the people interred in the Washington family cemetery?
Did you notice that I said, “mitochondrial DNA has been the chosen methodology” for identification?
The AFMES-AFDIL team is developing and refining multiple techniques that can be utilized to identify badly degraded remains of servicemen.
For example, in this case, there were only small bones, the DNA was severely degraded, and there was significant contamination.
If the mitochondrial DNA was a very common haplogroup, and was perhaps only partially recovered, they could eliminate several possible soldiers as matches, but they could not make a positive ID.
This case was just “problematic” enough to be useful, without being an unknown or unresolvable situation.
The family was involved and supportive. They knew who the candidate burials were in the cemetery and SWW contributed his own DNA for comparison.
SWW’s involvement provided two very important genetic benefits.
- First, SWW descended from Augustine Washington through the direct paternal line, so his Y-DNA should match that of the two Washington men in the burials.
- Secondly, SWW was related to the male burials in a short enough time period that he should match them both – one as his direct ancestor – his great-great-grandfather. The second burial was his great-great-grandfather’s brother. He should match his great-great-grandfather more closely than his great-great-grandfather’s brother.
| Individual |
Relationship to SWW |
Expected percent of DNA |
Expected cMs of DNA |
Relationship Degree with Dr. Samuel |
| Dr. Samuel Walter Washington |
Great-great-grandfather |
100 |
3500 |
|
| Christian Marie Washington married Richard Scott Blackburn Washington |
Great-grandmother |
50 |
1750 |
First |
| Samuel Walter Washington |
Grandfather |
25 |
875 |
Second |
| John Augustine Washington |
Father |
12.5 |
437.5 |
Third |
| SWW |
Present-day tester |
6.25 |
218.75 |
Fourth |
Lucy Payne would be SWW’s Fifth Degree relative, as would Dr. Samuel Walter Washington’s brother.
Full siblings share approximately 50% of the same DNA, so SWW would be expected to match the burial to whom he was more closely related with approximately twice as much autosomal DNA.
Therefore, using pairwise comparisons and kinship predictions, the team was able to discern which burial belonged to Dr. Samuel Walter Washington, because SWW matched that burial more closely.
But it turned out to be not quite that simple.
The Monkey Wrench
Relationships are classified as degree levels, as shown above. For example, children are first-degree relatives of their parents, siblings, and children. Genetic relationship levels are determined by comparing the DNA of two people and result in kinship predictions.
Normally, genealogists don’t think much about relationship degrees because we use the number of shared or overlapping centimorgans (cMs), and DNA testing companies provide kinship predictions.
However, because the AFMES-AFDIL team wasn’t working with the normal autosomal chip, they were only able to utilize a portion of the 95,000 locations, and they needed to “convert” SWWs results to compare to Dr. Samuel Washington and George Steptoe Washington Jr. They also needed to compensate for the fact that they were not able to obtain 100% of the 95,000 SNP locations on any of the burials. Recovered DNA ranged from 50%-85%
However, the burials matched SWW at one relationship degree level higher than expected.
Initially, Goran had asked me to review and work on expanding the genealogy of the Washington family, but now we had a new, very-interesting, wrinkle.
On a call, the team mentioned the disparity in the expected relationship level. I realized that the probable answer was that SWW was descended from Augustine Washington not just once, not twice, but three times, and we were seeing the genetic effects of pedigree collapse.
Those multiple relationships are beneficial when they provide one path to the Washington Y-DNA through a direct line to Augustine through his son, John Augustine, and another shorter path to Dr. Samuel Walter Washington for autosomal matching.
However, multiple relationship paths added complexity to autosomal relationship determination
There was yet a third avenue of descent to SWW through the father of Richard Scott Blackburn Washington, John Augustine Washington II.
In other words, there are three ways that SWW can and did inherit autosomal DNA from the Washington lineage, beginning with Augustine. Carrying extra autosomal DNA would affect the expected degree of relationship, potentially for SWW with both of the male Washington burials.
We needed a methodology to account for that.
Pedigree Collapse
I’m sure that the AFMES-AFDIL team didn’t view pedigree collapse as a benefit, at least not initially. They aren’t genealogists, so they really weren’t thinking about pedigree collapse in the same way genealogists do.
I’ve worked with pedigree collapse many times, but three separate events in the same line within a few generations was challenging in terms of getting the math right. It’s not obvious, and it’s not easy.
With pedigree collapse, it’s not just a simple matter of figuring out the expected percentage of DNA for all three relationships and adding them together because some of that DNA can be expected to be shared, which reduces the matching amount of DNA from the “add-three-together” number. So, the actual expected amount of shared DNA is someplace between the closest relationship, in this case, Dr. Samuel Walter Washington, and the additive result of all three relationships.
Plus, I couldn’t use cMs, so one hand was tied behind my back.
Therefore, we worked together to solve this puzzle.
My article, Pedigree Collapse and DNA – Plus an Easy-Peasy Shortcut is the result of my pedigree collapse calculations for this project – and how to make pedigree collapse easier for you to understand and account for.
It’s also the foundation of what I provided for the AFMES-AFDIL team, which integrated it into their protocol. Of course, when I published my Pedigree Collapse article, I had to remove anything that might have given anything away before the study and resulting paper was ready for publication.
Why the Monkey Wrench is Important
When dealing with unknown remains, we don’t have the luxury of already knowing who the family is and their potential position in the family.
The AFMES-AFDIL team wants to be able to utilize the techniques they are perfecting for the identification and repatriation of military remains as far back as WWII, 80 years ago. That means that those men would have been born nearly a century ago, and if a generation is roughly 20-25 years, the people available today to test may be as many generations removed from WWII veterans as SWW is from Dr. Samuel Walter Washington.
The repatriation team also won’t know if they are dealing with pedigree collapse until they see it. If a potential relationship comes back slightly differently than expected, they will know to consider either endogamy or pedigree collapse. Furthermore, tools that measure runs of homozygosity (ROH) can help inform them of either condition.
I’m glad this monkey wrench crept into the equation, and I was in the right place at the right time to help.
The Conversation
I joined this team someplace midway in the process, so I didn’t initially have the benefit of understanding why Courtney’s team was involved – that they hoped to refine their processes to begin utilizing autosomal DNA for repatriation.
I opined at one point that I was incredibly frustrated that this many years following the use of autosomal DNA for genealogy, the military was just now beginning to consider its use for repatriation, AND that they were not and had not been collecting autosomal DNA from family members of MIA/POW service members.
Courtney hopes this study will open that door sooner rather than later. As far as I’m concerned, next week would be great!
I was shocked that I had fallen into this opportunity, given that I have a POW/MIA family. member.
I’m a Gold Star Family Member
My first cousin, Robert Vernon Estes, Bobby, served in the Army in the Korean conflict. He was captured on November 30, 1950 in the horrific battle later known as “The Gauntlet.” He died on approximately January 31, 1951 in a POW camp someplace near Pugwon, Korea. He was only 19.
I am his namesake, and I also represent him as a Gold Star family member.
I’ve written about Bobby’s story, obtaining and unraveling his military records.
Bobby probably starved to death, as other members of his battalion did.
His mother died shortly after his capture, and he had no sisters to contribute mitochondrial DNA.
I’m the closest family member left now. We shared grandparents.
In July 2021, Bobby was honored by the State of Indiana. He served from White County. I was incredibly proud to be his representative family member.
When I accepted the invitation to assist the AFMES-AFDIL team with the Washington family burials, I had absolutely NO IDEA that their goal was to validate and extend this technology and these techniques to service member repatriation.
Bobby’s mother was adopted, so I have absolutely no ability to locate someone with Bobby’s mitochondrial DNA, which has frustrated me greatly for years. Therefore, if Bobby’s body were returned from North Korea today, his remains would remain unidentified and unclaimed. That possibility breaks my heart.
North Korea, “isn’t even answering the phone right now,” so the hope that Bobby will be returned to us in my lifetime fades a little with each passing day. That’s EXACTLY why it’s so important for the military to adopt and accept autosomal DNA from family members, even if they can’t utilize it today. My DNA and others can be archived for the future. Someday, Bobby and other servicemen may come back home.
Mitochondrial DNA alone couldn’t have solved the Washington mystery. There will be service members like Bobby who have no mitochondrial DNA sample waiting to be matched to them.
Just a few months before Goran asked me if I wanted to assist with a fun project, I had spoken with Bobby’s military representative, begging them to accept my autosomal DNA. No dice – at least not then.
Hopefully soon – very soon, so that we can begin to build the bank.
These men deserve to be identified. They gave their lives, their futures – that’s the least we can do for them.
The very least.
I’m so proud to be a part of this fantastic project. I’m incredibly grateful that Fate decided to put me in the right place at the right time, with the right combination of skills. I hope Courtney succeeds in pushing this door all the way open. It’s past time, and our team has proven beyond a doubt what can be accomplished. Our POW/MIA servicemen, servicewomen, and their families deserve it.
Thank you to my colleagues, Michael Sager and Goran Runfeldt at FamilyTreeDNA, Courtney Cavagnino, and the AFMES-AFDIL team.
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