A human skull, hidden between walls, was perhaps the last thing owners of a Batavia, Illinois, home expected to find during a 1978 renovation. Police investigated but, finding nothing, sent the unidentified remains to a university anthropology department. Scientists there determined the person was a woman in her mid-twenties and probably had died before 1900.

In 2021, the Batavia Police reopened the case and in 2023 they brought in Othram, a forensic DNA specialist laboratory, to sequence the DNA and identify potential relatives.

Eventually, the woman was identified as 17-year-old Esther Ann Granger, born in 1848. She had died 157 years earlier, in 1866, and was buried in Indiana before­‑—investigators think —someone exhumed and sold her remains, possibly for medical education.

The Granger case is remarkable because of the age of the DNA they were able to analyze, but the true value of forensic genomic analysis lies in identifying more recent cases involving missing persons and violent criminals. Othram’s website tells the stories of more than 500 men, women, and children who disappeared, and whose bodies were identified only because of forensic DNA. The technology is so powerful that the Bureau of Indian Affairs recently announced a joint project with Othram to help identify the remains of missing and murdered Native Americans and Alaskan natives.

“Forensic DNA analysis offers a way to solve cases that are truly unsolvable,” David Mittelman, PhD, CEO of Othram, says. “Using genomic tools in forensics is a force multiplier for investigations.”

SNPs and degraded DNA

“Forensics inputs are radically different than what you expect to see in the research or clinical setting,” Mittelman says. Unlike most clinical DNA sequencing settings, forensic DNA specialists are faced with samples that have been degraded by the elements or preservatives, making the fragments quite short. That, in turn, makes standard analyses using short tandem repeats (STRs) difficult, if not impossible. Instead, Othram’s analyses focus on single nucleotide polymorphisms (SNPs), which can be identified using much smaller fragments.

For context, a consumer DNA test collects between about 750 and 1,000 nanograms of DNA material. Othram accurately identified the perpetrator in a sexual assault case from a sample of only 0.12 nanograms.

Below 100 picograms (0.1 nanograms), allele dropout is likely, which means analysts may build a profile without the entirety of the genetic data needed to identify someone, rendering that profile less valuable.

Although Othram can successfully analyze small quantities and seriously degraded DNA, sometimes it must decline law enforcement requests. That’s usually because its lab hasn’t had success with that type or quality of sample. Rather than risk a limited sample, Mittelman says he would rather reject the sample until a more successful technique becomes available. “We err on the side of caution,” he stresses.

The company keeps a database of DNA types and quantities that it has rejected and is working—along with other DNA analysis companies—to develop techniques or technologies that, eventually, will allow it to take on those samples. “When we’ve validated a new method (in the research lab), then we can bring it to the forensics side,” Mittelman says, and reach out to the agencies that have experienced previously rejected samples.

Limited cross-fertilization

“Five years ago, there wasn’t much going on in this space,” Mittelman recalls. Even today, “There is relatively little cross-pollination between the genomics and forensics communities.” DNA analysis compares DNA profiles to known reference samples. Genomics analysis is broader, he explains. It uses whole genome sequencing to analyze the remains’ complete genetic makeup and may point investigators to research genealogy to help identify victims. Therefore, even when genomic tools exist, forensics teams may not have access to them, or the tools may still need to be optimized for forensics applications.

For Othram, that meant creating a purpose-built forensics genomics lab that “marries a forensics lab with a genomics lab powered by the Illumina NovaSeq platform.” The company also developed three new technologies: Forensics-Grade Genome Sequencing^® (FGGS^®), KinSNP^® for rapid kinship inferences in near or distant relatives, and the DNASolves^® Genetic Database. Adding genetic data supplied by individuals—often who have missing family members—makes definitive identification possible. Now the company identifies missing persons or criminal perpetrators nearly every day.

Today, Mittelman says, “We spend a lot of time working on DNA mixtures,” and other imperfect forensic inputs, and trying to find ways to not only make the identification process faster, but also more cost-effective, so more law enforcement agencies—“from the FBI to a local agency that doesn’t have a scientist”—can use it.

That said, “The important thing in forensics isn’t genotyping,” he continues. “It’s the ability to detect distant relatives, which allows you to piece together someone’s identity. The cross-pollination of these fields is a huge opportunity,” Mittelman says.

Forensic Genomics Pioneers

When Mittelman and his wife, Kristen Mittelman, PhD, chief development officer, founded Othram in 2018, DNA analysis and next-generation sequencing already had proven their value, but forensic genomic analysis was quite new. With the extra information it supplied, “law enforcement officers were excited but wondered whether this was science fiction. It’s science fact,” Mittelman told them, “but getting them to try it initially was a challenge.”

Now, with thousands of solved cases, it’s obvious forensic genomics works. His message today is that “it doesn’t just work for some cases. It can work for all cases.”

The issue today is funding. It’s important for law enforcement leaders to budget for forensic DNA analysis and for them to allow this technology to be used earlier in investigations, rather than reserving it as a last resort (especially when sample quantities are limited).

Othram works hand-in-hand with law enforcement, not only performing the biological work to build and analyze a profile and identify potential relatives, but also helping crowd-fund the work. “If you can find these pieces of data quickly, law
enforcement can find out who is responsible for the crime more quickly,” he says, “and minimize the chance they’ll prosecute the wrong person.”