Category Archives: DNA

April 13, 2017 by Liz Ahlberg Touchstone University of Illinois researchers developed a method to detect and map DNA methylation, which can be a sign of cancer, by threading the DNA through a tiny hole in a thin sheet of conductive material with a current running through it. Credit: Aditya Sarathy

Detecting cancer early, just as changes are beginning in DNA, could enhance diagnosis and treatment as well as further our understanding of the disease. A new study by University of Illinois researchers describes a method to detect, count and map tiny additions to DNA called methylations, which can be a warning sign of cancer, with unprecedented resolution.

The method threads DNA strands through a tiny hole, called a nanopore, in an atomically thin sheet of material with an electrical current running through it. The study was published in the inaugural issue of the journal npj 2D Materials and Applications, a new journal from Nature Press.

"One or a few methylations is not a big deal, but if there are many of them and they are packed close together, then it's bad," said study leader Jean-Pierre Leburton, a professor of electrical and computer engineering at Illinois. "DNA methylation is actually a starting process for cancer. So we want to detect how many of them there are and how close together they are. That can tell us at which stage the cancer is."

Other attempts at using nanopores to detect methylation have been limited in resolution. Researchers begin by punching a tiny hole in a flat sheet of material only one atom or molecule thick. The pore is submerged in a salt solution and an electrical current is applied to drive the DNA molecule through the pore. Dips in the current alert researchers that a methyl group is passing through. However, when two or three are close together, the pore interprets it as one signal, Leburton said.

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The Illinois group tried a slightly different approach. They applied a current directly to the conductive sheet surrounding the pore. Working with Klaus Schulten, a professor of physics at Illinois, Leburton's group at Illinois' Beckman Institute for Advanced Science and Technology used advanced computer simulations to test applying current to different flat materials, such as graphene and molybdenum disulfide, as methylated DNA was threaded through.

"Our simulations indicate that measuring the current through the membrane instead of just the solution around it is much more precise," Leburton said. "If you have two methylations close together, even only 10 base pairs away, you continue to see two dips and no overlapping. We also can map where they are on the strand, so we can see how many there are and where they are."

Leburton's group is working with collaborators to improve DNA threading, to cut down on noise in the electrical signal and to perform experiments to verify their simulations.

Explore further: Semiconductor membrane mimics biological behavior of ion channels

More information: Hu Qiu et al, Detection and mapping of DNA methylation with 2D material nanopores, npj 2D Materials and Applications (2017). DOI: 10.1038/s41699-017-0005-7

A semiconductor membrane designed by researchers at the University of Illinois could offer more flexibility and better electrical performance than biological membranes. Built from thin silicon layers doped with different ...

Gene-based personalized medicine has many possibilities for diagnosis and targeted therapy, but one big bottleneck: the expensive and time-consuming DNA-sequencing process.

Anyone who has been on a sail boat knows that tying a knot is the best way to secure a rope to a hook and prevent slippage. Similarly, knots in sewing threads prevent them slipping through two pieces of fabric. How, then, ...

Researchers from the National Institute of Standards and Technology (NIST) and collaborators have proposed a design for the first DNA sequencer based on an electronic nanosensor that can detect tiny motions as small as a ...

Collaborators from Mayo-Illinois Alliance for Technology Based Healthcare have developed a new, single molecule test for detecting methylated DNA. Methylationthe addition of a methyl group of molecules to a DNA strandis ...

Researchers at the National Institute of Standards and Technology (NIST) have simulated a new concept for rapid, accurate gene sequencing by pulling a DNA molecule through a tiny, chemically activated hole in graphenean ...

Detecting cancer early, just as changes are beginning in DNA, could enhance diagnosis and treatment as well as further our understanding of the disease. A new study by University of Illinois researchers describes a method ...

When you charge a battery, or when you use it, it's not just electricity but also matter that moves around inside. Ions, which are atoms or molecules that have an electric charge, travel from one of the battery's electrodes ...

Researchers have solved a problem hindering development of highly sensitive optical devices made of a material called graphene, an advance that could bring applications from imaging and displays to sensors and high-speed ...

Delivering life-saving drugs directly to the brain in a safe and effective way is a challenge for medical providers. One key reason: the blood-brain barrier, which protects the brain from tissue-specific drug delivery. Methods ...

Despite the many advances in portable electronic devices, one thing remains constant: the need to plug them into a wall socket to recharge. Now researchers, reporting in the journal ACS Nano, have developed a light-weight, ...

A new atomically thin material similar to graphene has been proven to be a promising new superconductive material.

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Nanopores could map small changes in DNA that signal big shifts in cancer - Phys.Org

INDIANAPOLIS Hoosiers arrested for any felony, starting in 2018, likely will have their DNA collected by cheek swab and their genetic record stored in a state police database.

State Sen. Erin Houchin, R-Salem, the sponsor of Senate Enrolled Act 322, said taking a DNA sample from every person arrested for a felony, instead of only those convicted of felonies, will help police solve more crimes similar to fingerprinting.

Opponents argued the DNA sampling is an illegal search, since police can use the DNA records for investigatory purposes, and not just identification.

"Our Constitution is pretty clear that you can't go and take things from people without a warrant to conduct an investigation. But here we're doing that," said state Sen. Mike Young, R-Indianapolis.

The legislation does contain a process for removing an individual's DNA from the database if the person is not convicted of a felony.

It passed the Senate, 36-13, and the House, 84-13.

Gov. Eric Holcomb now must decide whether to sign the measure into law.

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Indiana may create DNA record on every person arrested for felony ... - nwitimes.com

In Brief DNA protects itself from damage naturally, and scientists are hoping to gain insight into how the process works in order to achieve breakthrough treatments for cancer and other diseases that arise from the breakage and mutation of DNA. Simple Parameters, Complex Possibilities

DNA protects itself from damage naturally, and scientists are hoping to gain insight into how the process works. When DNA is bathed in ultraviolet light, it can eject a single proton from a hydrogen atom to rid itself of excess energy, ensuring other chemical bonds remain intact. This protective mechanism is called an excited state proton transfer, and it is the focus of new research by a team of scientists.

The researchers used the Linac Coherent Light Source (LCLS) at the SLAC National Accelerator Laboratory to generate X-ray laser pulses capable ofprobing the nitrogen molecule in the simple molecule 2-thiopyridone for quadrillionths of a second. The short period of time matters because when molecules are exposed to this kind of light they react incredibly quickly. The brightness of the light is equally important, because only very brilliant illumination renders these ultrafast changes visible to the researchers.

The researchers confirmed that optical light breaks nitrogen-hydrogen bonds, but the X-rays did not. This was important to the researchers because they wanted to confirm that any changes they were examining were related to the nitrogen atom or the area immediately surrounding it. The signature showing that the X-rays had been transformed into energy was the confirming resonance effect that they were looking for. On-resonance studies like this one allow researchers to clearly see how X-rays interact with samples because they amplify the signals for observation.

In future research, the team hopes to gain more insight into the greater class of photochemical reactions using the same approach to study more complex molecules. Knowing more about how molecules of all kinds protect themselves from radiation can help scientists achieve breakthrough treatments for cancer and other diseases that arise from the breakage and mutation of DNA.

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Scientists Witnessed DNA Protecting Itself From Radiation - Futurism

A state commission overseeing forensic science in New York is scheduled to take a key vote Wednesday that could pave the way for local law enforcement to begin using this summer the emerging technique of familial DNA searching to solve homicide and sex crimes.

The vote by the New York State Commission on Forensic Science is slated for Wednesday morning. The consent of at least seven members of the 14-person body is needed to approve regulations governing the use of the technique by DNA labs in the state, said a spokeswoman for the state Division of Criminal Justice Services.

A special DNA subcommittee last month approved the measure by a vote of 6-0, with one member absent. If the full commission approves the regulations, they would be published in the State Register and the public would have 45 days to comment before they take effect, DCJS spokeswoman Janine Kava said.

Familial searching is in use in 10 states, including California and Colorado. Law enforcement officials in Britain and the Netherlands also use it.

Law enforcement officials in New York City, including NYPD Commissioner James ONeill and Queens District Attorney Richard Brown, pushed for the procedure after a story last November in Newsday highlighted its potential use in the case of slain Howard Beach jogger Karina Vetrano.

Although the Vetrano probe led to an arrest using conventional methods, the family still supports familial searching.

This has to be done for other people, Vetranos father, Philip, told Newsday recently.

The searching method is called upon in cases where an unknown crime scene DNA sample as occurred in the Vetrano case has failed to match any genetic profiles in the state databases. The unknown sample is put through a two-step process that uses special software to produce a list of persons in the database who are likely close relatives of the person whose DNA was left at the crime scene. In the second step, a lineage test is done, usually involving the Y chromosome, to confirm the relationship.

Armed with the test results, police would question the identified relatives of the person under scrutiny to try and locate and obtain a DNA sample from the person of interest. In about 36 percent of cases the investigations lead to the identification of a suspect, according to a 2015 report by the National Institute of Justice.

Under the New York proposal, crimes for which the technique can be used include homicide, first-degree rape, sexual assault, arson, first-degree kidnapping and crimes involving a significant threat to public safety. Strict privacy controls are required.

Civil libertarians and some defense attorneys have criticized the technique. At a February public hearing, Erin Murphy, a professor at New York University School of Law, said it can put relatives of somebody with a criminal record unfairly under suspicion as a possible suspect simply because of a blood relationship.

Murphy also said that there was racial bias at play since people of color are convicted in numbers proportionately greater than their population sizes and have their DNA on file.

But proponents stress that the technique is race-neutral, can clear innocent people and that people of color are overrepresented in the population of both crime victims and suspects. A police source said of the 11 unsolved homicides in New York in 2016 with unknown DNA samples, nine involved black victims, one Hispanic and one white.

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State panel to consider new DNA technique to ID crime suspects ... - Newsday

Millions of years ago, our ancestors bodies may have waged an epic battle against a virus. Through a sneaky DNA swap, the hosts would have gotten the upper hand, and turned the virus defenses against themselves. Researchers think theyve unearthed the whole ordeal, left like a Dead Sea scroll in our genes.

We know very little about ancient and extinct viruses, since they (mostly) cant form real fossils. But retroviruses, the type of viruses that include HIV, plug their own genetic material into the cells they infect. If a retrovirus infects cells that get passed to offspring, then we can potentially see exactly where and when it was around by looking at ancient and modern DNA. A team of researchers from Rockefeller University did just that, and found what looked like a cunning act of bodily revenge where our extinct hominid cousins used a viruses DNA against itself.

So the whole story is very exciting to me (with a little bit of bias of course since it was my PhD thesis work), first author Daniel Blanco Melo told Gizmodo in an email. From one side we were able to reconstruct a very old [viral] gene from destroyed sequences in primate genomes. And on the other hand we encounter that our ancestors were able to take some information of this virus and used it to drive its own extinction.

Viral archaeology is forensic game. The researchers were interested in studying whether there were any examples in history of hosts evolving protection against viruses, and came across HERV-T, a virus that probably left its genetic mark in germline cells, those that get passed from generation to generation including fetal cells, sperm cells and egg cells. That means the researchers were able to look at the DNA of ancient hominids to see whether or not they had HERV, checking out more and more distant cousins until they found one without the viral DNA. The number of years ago that an infected cousin and our ancestors split off on the evolutionary tree tells us about how old the virus is.

The researchers also investigated how HERV-T DNA mutated over time, by comparing data previously collected from several old world monkey species genomes, like gibbons and bonobos. Like trying to get two different cultures to remember a myth over a very long period of time, minor details change, and differences accumulate between species. The team knew the approximate DNA mutation rates in different species, and used the differences to approximate age of the virus.

HERV-T probably emerged between 30 and 40 million years ago, embedding its DNA into the cells of our ancestors. But where is it today? Thats where our cells may have performed a cunning evolutionary act. Based on their primate analysis, the researchers reconstructed the protein described by the HERV-T gene remnants in the DNA of modern humans. This ancient protein embedded itself in the envelope that encapsulated the virus DNA. The host used that protein to evolve a derivative of it that can essentially block the receptor from the cell surface, Paul D Bieniasz, the papers principle investigator from The Rockefeller University in New York, told Gizmodo, thereby inhibiting the infection by the replicating virus.

Basically, the viral intruder had a copy of the key to the house, but left it inside. Armed with that knowledge, our ancestors cells changed all the locks. Or, more accurately, they got rid of the doorknobs. The primates with the viral gene would have won the survival-of-the-fittest game, and the virus probably went extinct around ten million years ago. However, it is important to note that one anonymous reviewer of the paper, published today in the journal eLife, had some concerns with the evidence that the hsaHTenv ORF, the genetic sequence coding the resurrected protein, was maintained due to selective pressure. The researcher didnt think the evidence that natural selection caused us to continue producing the protein was convincing.

Despite this caveat, the researchers findings could be a first in viral archaeology. Weve known its possible for hosts to co-opt envelope proteins to protect themselves from retrovirus infection, but this is the first time its been demonstrated in hominids, our direct ancestors, said Bieniasz. Its also the first time an extinct virus protein has been resurrected.

Its important to note that this is just one study about one protein, and a many million year look into the past requires some degree of speculationwe werent around to observe the virus or its effects, after all.

The researchers think they might be able to use the protein in cancer research. They think they could make their own viruses stuffed with the ancient viral protein to target and kill cancer cells. Its unlikely that wed be able to cure ourselves of other retroviruses, though, like HIV, since most of them dont infect the germline cellswe wouldnt be able to pass resistance on to our children, or get all of our cells to develop resistance.

Regardless, just think, your DNA essentially contains ancient scrolls with evolutionary epics waiting to be read.

[eLife]

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Scientists Unearth Ancient DNA Depicting a Battle Between Viruses ... - Gizmodo

April 11, 2017 In the original CRISPR-Cas9 technique (top), the guide RNA (green) binds to the target DNA and the cleavage enzyme Cas9 (scissors) cuts out a small DNA sequence (red). A modified version of Cas9 called nCas9 (bottom) is different as it cuts off only one strand of DNA and the cytidine deaminase (pink) transforms a single cytosine (C) into uracil (U). Uracil (U) is then converted to thymine (T) by DNA replication. Credit: IBS

Researchers at the Institute of Basic Science (IBS) proved the accuracy of a recently developed gene editing method. This works as "DNA scissors" designed to identify and substitute just one nucleotide among the 3 billion. "It is the first time that the accuracy of this base editor has been verified at the whole genome level," explains KIM Jin-Soo, leading author of the study. Published in Nature Biotechnology, this validation will help to expand the use of this method in agriculture, livestock, and gene therapy.

Rapid advances in gene editing tools have created excitement in the biology community. The primary third-generation DNA editing technology is CRISPRa tool that is quicker and cheaper than its predecessors. By cutting out a small DNA sequence, CRISPR-Cas9 and CRISPR-Cpf1 are used to silence or reduce the expression of faulty genes. However, last year, biologists discovered a new base editor method that does not cause random DNA deletions and insertions, but instead replaces only one DNA base. These types of gene corrections are critical, as several diseases are caused by the misspelling of one of the four basic components of DNA; adenine (A), cytosine (C), guanine (G), and thymine (T). Single-nucleotide errors in DNA are referred to as point mutations. Examples of conditions caused by point mutations include cystic fibrosis, sickle cell anemia and color blindness.

Unlike the existing technologies, the base editor method consists of a variation of CRISPR-Cas9 (nCas9, nickase) fused with another enzyme called cytosine deaminase, which replaces the DNA component C with T. The scissors are directed to the correct position on the DNA by a guide RNA. However, until recently, it was not known whether the base editor was working only in the area of the faulty gene or if it was unnecessarily substituting Cs in off-target areas.

Just one month after reporting the first successful base editing in animals in Nature Biotechnology to modify a single nucleotide in dystrophin and tyrosinase genes, the same team demonstrated the accuracy of this method at the genome scale.

In order to identify the soundness of the method, IBS researchers modified the error-checking technique known as Digenome-seq, in order to adapt it to the base editor method. Digenome-seq was used and validated last year, when the team analyzed the accuracy of CRISPR-Cpf1 and Cas9. IBS researchers also improved the computer program Digenome 2.0 to identify off-targets more comprehensively and compared different guide RNAs to find the one that reduces malfunctions and increases specificity.

Using this technique, the team found the base editing technique to be even more accurate than the current third-generation CRISPR-Cas9. The base editing technique induced C-to-T conversions in multiple sites in the human genome, while CRISPR-Cas9 caused cleavages in multiple sites, meaning that the new base editor technique makes fewer off-target changes. "Therefore, it is expected that these base editors will be used as widely as the popular CRISPR technology," says KIM.

Explore further: First CRISPR single-nucleotide edited transgenic mice

More information: Genome-wide target specificities of CRISPR RNA-guided programmable deaminases, Nature Biotechnology (2017). nature.com/articles/doi:10.1038/nbt.3852

Cystic fibrosis, sickle cell anemia, Huntington's disease and phenylketonuria are all examples of disorders caused by the mutation of a single nucleotide, a building block of DNA. The human DNA consists of approximately 3 ...

A team from the Center for Genome Engineering, within the Institute for Basic Research (IBS), succeeded in editing two genes that contribute to the fat contents of soybean oil using the new CRISPR-Cpf1 technology: an alternative ...

(Phys.org)A team of researchers affiliated with Harvard University and the Howard Hughes Medical Institute has announced the development of a gene-editing system that improves on the performance of CRISPR/Cas9 by allowing ...

(Phys.org)A team of researchers with members from several institutions in Japan has developed a new way to edit genes that involves cutting just one strand of DNA rather than both of them, as is normal for CRISPR-Cas9. ...

CRISPR-Cas9's popularity continues to grow ever since its first use in genome editing in January, 2013. What makes CRISPR-Cas9 so remarkable is its astonishing efficiency and availability; relatively speaking, it is easy ...

Researchers from Memorial Sloan Kettering Cancer Center (MSK) have harnessed the power of CRISPR/Cas9 to create more-potent chimeric antigen receptor (CAR) T cells that enhance tumor rejection in mice. The unexpected findings, ...

A University of Wyoming weed scientistfrustrated with the noise surrounding genetically modified organisms and glyphosate useanalyzed data to see for himself if biotech adoption has had a negative or positive effect ...

How can you tell if an individual is expressing sexual interest? With males, it's usually quite obvious and can be anything from lavish theatrical displays of song and dance to downright relentless insistence. Females, on ...

Researchers at the University of Alberta have demystified the way that polar bears search for their typical prey of ringed seals. The answer, it turns out, is simple: they follow their nose using the power of wind.

Asian elephants are able to recognise their bodies as obstacles to success in problem-solving, further strengthening evidence of their intelligence and self-awareness, according to a new study from the University of Cambridge.

Millions of years before humans discovered agriculture, vast farming systems were thriving beneath the surface of the Earth. The subterranean farms, which produced various types of fungi, were cultivated and maintained by ...

One of the largest colonies of gentoo penguins in Antarctica was decimated by volcanic eruptions several times during the last 7,000 years according to a new study. An international team of researchers, led by British Antarctic ...

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Accurate DNA misspelling correction method - Phys.Org

Environmental DNA helps protect great crested newts -- ScienceDaily Science Daily Research has revealed how tiny amounts of DNA (eDNA) released into water by great crested newts can be used to monitor the species. This can bring benefits ... and more »

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Environmental DNA helps protect great crested newts -- ScienceDaily - Science Daily

The Daily Advertiser archives Published 3:24 p.m. CT April 11, 2017 | Updated 16 hours ago

Sirens(Photo: Getty images)

This story was originally published Aug. 6, 2011. Aaron Richards was convicted of first-degree murder in this case.

Confusing new DNA evidence in a first-degree murder case could possibly help the accused killer go free.

Test results of DNA found on the pants of Tim Falgout, the pizza delivery man who was killed in Youngsville in March 2010, indicate the involvement of a man who died days before Falgout was killed.

"We always said from Day 1 there was a problem with the DNA evidence," said Harold Register, the court-appointed attorney for Aaron Richards, who is charged with the first-degree murder of 50-year-old Falgout.

"The fact that we've gotten evidence that another person's DNA was mixed in shows it's a classic case of poor police work," Register said.

Richards, 35, of Kaplan, pleaded not guilty after police tied his DNA to the knife used to kill Falgout. The new DNA, however, points to a man named Thomas Evans.

Evans, 26, of Boyce, was killed one week before Falgout was murdered. Evans' killer killed himself during a police standoff.

Court documents show a Lafayette law enforcement officer contacted Rapides Parish about the possible connection but ultimately dismissed the idea.

"I have no idea how (Evans') DNA got there," Register said. "That's a mystery. It's pretty interesting, and it puts a twist in the case."

Falgout, who was delivering a pizza on the 300 block of Canard Street in Youngsville, was found dead March 29, 2010, in front of a customer's home. Falgout's wallet was missing, and officials cited robbery as the motive.

Police also arrested a second man in the case for his alleged role as the getaway driver.

Police charged Marcus Feast with principal to first-degree murder and say he was driving a gold Mercedes, which was used to follow Falgout during his pizza delivery and flee the scene after the robbery.

Richards, who was arrested Aug. 5, 2010, remains in the Lafayette Parish Correctional Center, where he is being held without bond.

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From the archives: DNA complicates murder case - The Daily Advertiser

New Method Offers Promise of DNA-Based Vaccines in Pill Form P&T Community Researchers at the University of NebraskaLincoln have demonstrated that nesting specialized nanoparticles inside microparticles could protect engineered genes or virus-derived DNA against the rigors of the stomach and ensure safe passage to the ... and more »

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New Method Offers Promise of DNA-Based Vaccines in Pill Form - P&T Community

Darlene Krashoc was slain 30 years ago, and now authorities say they know what her murderer looks like. Its given her parents hopeand thats what forensic experts fear.

DENVERPaul Krashoc waited 30 years to come face to face with his daughters killer, but not like this.

Staring back at him the dining room table were two computer-generated images of the man the U.S. Army suggests murdered Specialist Darlene Krashoc on March 17, 1987, outside Fort Carson, Colorado. One image showed what he might have looked like back then, in his twenties, and the other shows what he might look like today, in his fifties.

This new spectrum of info, it hit us quickly. This opens up new doors for us, Krashoc told The Daily Beast. He acknowledges, though, that after 30 years, he and his wife Betty Lou have been down this road before.

Our daughter has been dead longer than she lived, Betty Lou said. She said she keeps a picture of Darlene in her Bible along with the printout of those two faces.

The Army released the images last month after sending DNA data from 27 pieces of evidence to a private lab called Parabon to undergo a type of analysis called phenotyping. Phenotyping uses unidentified DNA to predict appearance and ancestry. In its press release, the Army heralded this new development as state of the art science. Its a statement that has some DNA scientists and law enforcement experts fuming.

Phenotyping is not state of art. Its not the state of anything, says retired Denver homicide detective Jon Priest. He is concerned that phenotyping gives false hope to victims families who are desperate for answers.

The new images have given Paul Krashoc new energy. He has always suspected that a fellow soldier killed Darlene, and now the phenotype has him convinced.

This picture. If you look at the haircut, it indicates military, he said.

Krashocs comment is exactly what worries scientists who have been in the field of forensic DNA for years. They say the haircut on the rendering is only a suggestion to help round out the picture, not science.

Its giving the family unrealistic expectations, says Dr. Richard Spritz, director of Human Medical Genetics and Genomics at the University of Colorado School of Medicine. Do I think its likely having this technology will get them to a perpetrator? No. Because the reliability of this technology has never been subject to critical tests. The likely reliability is low.

Priest echoed the concern.

This is one of those things that is not proven, supportable, nor accepted, Priest said, adding hes not against trying new technology. The science may get there someday, but it aint there now.

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Ellen Greytak of Parabon Labs said she understands that people are skeptical, but adds that they might change their minds if they see a presentation and realize how strong our results are.

Parabon, which unveiled the technology just over two years ago, has never had a dissatisfied client, Greytak says. Parabons phenotyping process isnt meant to produce an exact replica of a persons face, but rather an image that strongly resembles the person the unidentified DNA belongs to.

It helps narrow down suspects gives law enforcement the ability to eliminate them, Greytak explained.

On its website, Parabon gives law enforcement a special offer of a free online demonstration. Also offered are testimonies from various investigation-related entities who swear by the technology, including Jose Morales, a detective with the Costa Mesa, California police department.

We had a cold case for 20 years. We gave up hope. These kinds of cases, they weigh on us, Morales tells The Daily Beast. His investigation into the random rape and murder of a photography student was solved when the Parabon rendering, plus fingerprints left at the crime scene, linked the killing to a man who was arrested in Mexico and is awaiting extradition.

Still, Spritz takes issue with how Parabon describes its product as a Snapshot Forensic DNA Phenotyping System, which accurately predicts genetic ancestry, eye color, hair color, skin color, freckling, and face shape

Spritz agrees that DNA can determine eye, hair, and skin color, but he stops short at face shape.

We do not have the knowledge to predict a face based on DNA, Spritz said.

Greytak says it costs $3,600 for Parabon to analyze the DNA data and come up with its snapshot. But Greytak admits that out of the 100 cases theyve been involved with, only one has been resolved in the courts.

Its not probable cause, and thats the real problem with it, says Priest. If I find somebody it looks like, I cant arrest the guy!

The Colorado Springs police, who have been investigating Krashocs murder in tandem with the Army since the beginning, also notes that the pictures are not evidence.

We need to be careful, said Lt. Howard Black. We would never bring someone in just based on a picture. But he said the renderings have brought people with information out of the woodwork. They do help. If an individual looks at them, the images can help bring back conversations from 30 years ago.

The Army is also offering a $10,000 reward to anyone who can help find a murderer who has eluded them through three decades that saw the cold case opened, closed, and re-opened again in 2004 and in 2011 as DNA technology advanced.

Though years of fruitless leads have put a chill on the military whodunnit, the brutality of the murder of the beautiful young soldier stunned the town an hour and a half south of Denver.

Veteran Colorado Springs Gazetteeditor Tom Roeder says he has never seen anything like it in 20 years of covering the Army at Fort Carson.

Weve had murders as a result of domestic abuse, but never anything like this. Roeder adds, We see from other cold cases at Fort Carson that the C.I.D. [U.S. Army Criminal Investigation Command] usually closes the door and walks away. When the trail goes cold they shove it in the file thats been their tendency. Still, someone knows something about this case.

Krashoc had been out at an off-base nightclub called Shuffles on the eve of St. Patricks Day, 1987, drinking with friends who worked with her in the 73rd Wheeled Maintenance. Her parents say a friend later told them she got mad when she couldnt get a cigarette from one of them, and put up such a scene about it, they left her there.

Just before dawn police found Krashocs body dumped in an alley behind a Korean restaurant. They think the 20-year-old Army specialists body was brought there from somewhere else, because of the time it must have taken to torture her. The autopsy report even suggested she may have been thrown out of a moving car. The woman friends had nicknamed Krash had been sexually assaulted, beaten, bitten, and, strangled with a coat hanger.

That clue led her parents to believe the killer was a soldier named Jeffrey Newsome, who was stationed at Fort Carson and accused by two women of strangling them with a coat hanger before he was convicted of murder in Alabama.

Newsome has been eliminated as a suspect, according to police though they wont divulge the reason. Lt. Black did say that there has never been a match with the DNA found on Krashocs body through CODIS, a database that compares samples found from crime scenes to the DNA of convicted felons and sometimes, to arrestees. Since Newsome is in prison for life, that means his DNA did not come up as a hit.

Because the Krashocs anticipated that they may have to exhume Darlenes body for more forensic evidence, they buried her in a civilian cemetery instead of in a military one where its much harder to get an exhumation done.

The 30-year stress of never finding Darlenes killer has been too much for their relationship with people who loved her the most. They say that they no longer speak to their other daughter, Darlenes sister, Rhonda, and the same goes for Darlenes best friend who was stationed at Fort Carson and even roomed with her.

Long years of searching with no answers gnaws at everyone involved, and so does a statement Darlene made the last time Betty Lou spoke with her on the phone.

Theres something going on at Fort Carson, Darlene told her mom. But the young woman would not reveal anything more.

I asked her, What the hells going on out there? and she just said, I cant tell you right now. The weird thing is, just a week before that phone call, Darlene was talking about re-enlisting.

Betty Lou said she was afraid her daughter was thinking about going AWOL, but instead, says her husband, A week later, the guy was at the door in uniform to tell us she was dead.

The Krashocs do not believe that the Army did all that they could have when memories were fresh. They recall that early on during the investigation, they walked into the Office of Command at Fort Carson, and they were turned away.

They didnt want publicity, says Paul, a veteran Army deep sea diver. We were shaken up, but thats when we decided to become a pain in everyones rear end.

Through the years, the Krashocs have done their share of amateur sleuthing. Betty Lou has cardboard boxes in their home office filled with newspaper clippings of similar murders which happened all over the world.

I look them up, call the police in charge, and let them know about Darlene.

The snapshot DNA sketches and Darlenes picture will stay in Betty Lous Bible. Theyve read the comments on social media about how the new computer images are a long shot. And they know their hopes could be sunk again. Still

We could be getting fed a lot of bull, says Paul. Obviously we dont know how this will turn out. But what else can we do?

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Is Sketching a Killers Face From DNA Science or a Scam? - Daily Beast