Apr 18 2016

Autosomal and Y-STR Analysis of Degraded DNA from 120-Year-Old Skeletal Remains

Interesting CasesMediaMassively Parallel SequencingForensic

Angie Ambers of the Institute of Applied Genetics (UNTHSC) describes how she processes skeletal remains to determine an individual’s identity, and how she identified a Confederate guerilla scout from the American Civil War using this technology.





My name is Doctor Angie Ambers, and I work for the Institute of Applied Genetics, which is an institute at the University of North Texas Health Science Center in Fort Worth, Texas. I primarily work on characterization and DNA analysis of historical and archaeological skeletal remains, although I do do work in developing improved methods for skeletal remains analysis.

This poster is a really interesting case. This was an individual named Ezekiel Harper, and he was a pretty famous (or infamous) Confederate guerilla scout from the American Civil War. He actually survived the war and became a prominent, wealthy doctor in the West Virginia area. He was subsequently (after the Civil War) beaten to death and murdered in a home invasion.

He had amassed quite a bit of land. He owned 4,000 acres, which back in the 1800s was quite a bit. When he was murdered, he was single – he never married. There was a small boy that allegedly was the son of his affair with his Native American maid. So, it’s kind of a Thomas Jeffersonish type case that was kind of scandalous during that time period. After he was murdered, of course, the maid was sent away and the little boy was sent to the county farm, which essentially was an orphanage at that time.

The little boy ended up growing up and having seven children of his own. He grew up telling all the locals that he was related to the infamous Ezekiel Harper. His family, and his children over the generations, followed the development of the DNA field and when they thought the time was right, they petitioned the state of West Virginia to exhume Ezekiel Harper’s remains to do testing to see if there was a familial link between the alleged son and Ezekiel Harper.

We did Y-STR Y-Chromosome testing based on the family members that were still living that were available to give reference samples. So, we did establish a familial link between the alleged son and Ezekiel Harper.

The children of this individual, his name is Earl Maxwell (the alleged son of Ezekiel Harper), he was kind of ridiculed. He grew up in an orphanage and he was telling everybody he was the son of this famous guerilla scout, this famous Confederate Civil War soldier, and everybody kinda thought he was crazy, so this kind of vindicates the family, and gives them that “Ok, we’ve had DNA testing done and it does establish a link between our father, or our grandfather, and Ezekiel Harper.” It’s a little bit of vindication for him as well as the potential for regaining that land.

Working on skeletal remains is particularly challenging. Processing the bone samples takes quite a bit longer than your standard samples like blood or semen or saliva. Typically the process involved in that is  we get the skeletal remains in, we take a look at what skeletal elements we have available for testing, kind of decide which bones would be most appropriate, or might give us the best chance of recovering DNA. And we literally have to, it’s a destructive technique, we have to saw the bones into small sections and then we put these sections into sterile vials and submerge them in liquid nitrogen. With that liquid nitrogen, we grind the bone into a fine powder. It almost comes out to be similar to a baby powder format, and that’s how we extract the DNA from the samples.

That processing takes quite a bit of time, as well as all of the contamination prevention techniques and procedures that you have to go through to prep all of the materials just to work with those bones.

With skeletal remains, because the DNA tends to be pretty damaged sometimes, depending on the passage of time and the environmental conditions to which those skeletal remains were exposed, often what we will do is sample multiple different regions of the skeleton, and/or we’ll do replicate testing. So, sometimes what you’ll end up getting is a partial DNA profile from one sample and then you’ll test another sample that you’ll also get a partial profile, and you’re hoping that the partial profile that you got from one bone section is a little bit different, different markers, different results with different markers, from this other bone section. If you put enough of those partial DNA profiles together, the goal is to hopefully put together a consensus DNA profile.