Ancient DNA Methods Improve Forensic DNA Profiling of Korean War and World War II Unknowns

The integration of massively parallel sequencing (MPS) technology into forensic casework has been of particular benefit to the identification of unknown military service members. However, highly degraded or chemically treated skeletal remains often fail to provide usable DNA profiles, even with sensitive mitochondrial (mt) DNA capture and MPS methods. In parallel, the ancient DNA field has developed workflows specifically for degraded DNA, resulting in the successful recovery of nuclear DNA and mtDNA from skeletal remains as well as sediment over 100,000 years old. In this study we use a set of disinterred skeletal remains from the Korean War and World War II to test if ancient DNA extraction and library preparation methods improve forensic DNA profiling. We identified an ancient DNA extraction protocol that resulted in the recovery of significantly more human mtDNA fragments than protocols previously used in casework. In addition, utilizing single-stranded rather than double-stranded library preparation resulted in increased attainment of reportable mtDNA profiles. This study emphasizes that the combination of ancient DNA extraction and library preparation methods evaluated here increases the success rate of DNA profiling, and likelihood of identifying historical remains.

DNA Profiling from Dentine and It’s Role in Signifying Individuality

Forensic dentistry is the field which is a combination of forensic and dentistry. In this we go for principles used in dentistry or dentistry forthe use in judiciary. It is one field inwhich we collectthe evidence related to dentistry and analyze them for the purpose of investigation. In a crime scene or a disaster situation a lot of destruction occurs. It becomes important to determine the identity of an individual and we can say it is the identity of the deceased. This study is being conducted based on previous research and literature presented by various research scholars. When it comes to a crime scene where we do not get the victim or culprit but we find the dentalevidence and also in cases of mass disasters, catastrophic events, industrial disasters where identification is necessary if we found the dental evidence, they can help a lot in determining age, gender partial identity can be known through this evidence. Many methods are there to determine identity through dental evidence and also apart from dental evidence we have orthometric methods but dental evidence is found to be more cheap, easy, fast methods. In this we have discussed how DNA is extracted from the teeth and it's further processing so that it will help in establishing someone's identity. Keywords: Dentistry, Forensic Dentistry, Dental Evidence, Extraction, DNA Profiling etc

Role of Forensic Anthropology in Disaster Victim Identification (DVI)

DNA profiling is one of the most dependable and well-organized methods for recognizing bodies or losing body parts in disaster victim identification (DVI). This necessitates the collection of a post-mortem DNA sample and an antemortem DNA sample from the alleged victim or a biological related people. Collecting an acceptable ante mortem sample is usually effortless, but because of the varying degree of preservation of the human remains after any disaster and very high risk of cross-contamination of DNA, obtaining an adequate standard post mortem sample under cold DVI conditions is difficult. Various post mortem DNA samples from a deceased person in DVI can be obtained from muscle, bone including femur and ribs, teeth, and bone marrow with the slightest possibility of contamination. DVI (disaster victim identification) has been used to identify deceased people in various famous disasters like the 9/11 attack of the terrorist group al-Qaeda against the United States, Malaysia Airlines Flight 17 from Amsterdam to Kuala Lumpur that was shot down on 17 July 2014 while flying over eastern Ukraine. All 283 passengers and 15 crew members were killed; the 26/11 attack on Mumbai in 2008 led by terrorist organizations caused 166 deaths, excluding nine terrorists. According to Interpol protocol, four steps for identification are given importance: 1 – Site examination, which lasts for days to weeks. 2 – Post-mortem data include fingerprints, odontology, DNA profiling and physical indication. 3 – Ante-mortem data collected from victim houses. 4 – Reconciliation where specialists identify the victim from the data collected.

Forensic DNA Profiling of Bacterial Communities in Soil

<p>Soil is frequently encountered as trace evidence in forensic science case-work, but because of the limitations of current analytical techniques, this evidence is rarely utilised. A technique has been developed that allows comparisons of soil samples to be made, based on molecular analysis of the bacterial communities living in the soil. This project assesses the practicality of using this technique, known as 16S rDNA T-RFLP community profiling, for forensic soil analysis, by refining the basic methodology and performing a preliminary evaluation of its reproducibility and utility. Initial difficulties associated with generating profiles from soil samples have been overcome through methodology improvement, and the technique has been found to be effective for generating simple, visual profiles that clearly demonstrate differences between soil samples. Soil bacterial community DNA profiling is likely to be a powerful yet simple forensic tool, providing the ability to routinely use soil as associative evidence. The potential for using the same technology to develop a time since death or post mortem interval (PMI) estimation tool was also investigated. This study monitored the changes in the soil bacterial community beneath decomposing human cadavers and pig carcasses and showed that community change is dynamic and progressive. These changes are caused by fluctuations in specific bacterial species populations that are able to utilise organic breakdown products released from the body over time. Release of the body’s natural microflora into the underlying soil may also contribute to an altered bacterial community. This project has demonstrated that the soil microbial community clearly changes over the course of decomposition, and potential exists for development of a PMI estimation tool based on soil bacterial community succession.</p>

Forensic DNA Profiling of Bacterial Communities in Soil

<p>Soil is frequently encountered as trace evidence in forensic science case-work, but because of the limitations of current analytical techniques, this evidence is rarely utilised. A technique has been developed that allows comparisons of soil samples to be made, based on molecular analysis of the bacterial communities living in the soil. This project assesses the practicality of using this technique, known as 16S rDNA T-RFLP community profiling, for forensic soil analysis, by refining the basic methodology and performing a preliminary evaluation of its reproducibility and utility. Initial difficulties associated with generating profiles from soil samples have been overcome through methodology improvement, and the technique has been found to be effective for generating simple, visual profiles that clearly demonstrate differences between soil samples. Soil bacterial community DNA profiling is likely to be a powerful yet simple forensic tool, providing the ability to routinely use soil as associative evidence. The potential for using the same technology to develop a time since death or post mortem interval (PMI) estimation tool was also investigated. This study monitored the changes in the soil bacterial community beneath decomposing human cadavers and pig carcasses and showed that community change is dynamic and progressive. These changes are caused by fluctuations in specific bacterial species populations that are able to utilise organic breakdown products released from the body over time. Release of the body’s natural microflora into the underlying soil may also contribute to an altered bacterial community. This project has demonstrated that the soil microbial community clearly changes over the course of decomposition, and potential exists for development of a PMI estimation tool based on soil bacterial community succession.</p>

PEMERIKSAAN DNA SENTUHAN DARI BARANG BUKTI DENGAN PERBEDAAN JENIS BAHAN

Analysis of Touch DNA on forensic laboratory has been a favorite approach to identify a person. Every investigator demand the identity of whom the perpetrator that commit the crime, that leaved their DNA on the evidence. Many factors affect touch DNA, one of these is the substrate of the evidence. Common evidences that often examined in forensic lab are firearms, knife, swords, clothes, and switch bomb. To collect the cell on the evidence we use tapelift method using the duct tape. PrepFilerTM BTA Extraction Kit used to extract the DNA from the duct tape, followed by Quantifiler® Duo. For profiling the DNA we use GlobalFilerTM and fragment analyzed on ABI 3500 Genetic Analyzer followed by GeneMapper ID.X. V.1.4. Based on our analysis, DNA from fabric substrate has the higher percentage of success DNA profiling. The success DNA profiling rate of fabric and plastic substrate is 100%, and 0% for wood substrate. According to recent researches, smooth substrate, like plastic and glass, has higher percentage to get full profile than rough substrate, like woods. But on the fabric, they found has much higher percentage than smooth substrate. This can be due to the absorption ability of the fabric to obtain more cells

Primer Binding Site (PBS) Profiling of Genetic Diversity of Natural Populations of Endemic Species Allium ledebourianum Schult.

Endemic species are especially vulnerable to biodiversity loss caused by isolation or habitat specificity, small population size, and anthropogenic factors. Endemic species biodiversity analysis has a critically important global value for the development of conservation strategies. The rare onion Allium ledebourianum is a narrow-lined endemic species, with natural populations located in the extreme climatic conditions of the Kazakh Altai. A. ledebourianum populations are decreasing everywhere due to anthropogenic impact, and therefore, this species requires preservation and protection. Conservation of this rare species is associated with monitoring studies to investigate the genetic diversity of natural populations. Fundamental components of eukaryote genome include multiple classes of interspersed repeats. Various PCR-based DNA fingerprinting methods are used to detect chromosomal changes related to recombination processes of these interspersed elements. These methods are based on interspersed repeat sequences and are an effective approach for assessing the biological diversity of plants and their variability. We applied DNA profiling approaches based on conservative sequences of interspersed repeats to assess the genetic diversity of natural A. ledebourianum populations located in the territory of Kazakhstan Altai. The analysis of natural A. ledebourianum populations, carried out using the DNA profiling approach, allowed the effective differentiation of the populations and assessment of their genetic diversity. We used conservative sequences of tRNA primer binding sites (PBS) of the long-terminal repeat (LTR) retrotransposons as PCR primers. Amplification using the three most effective PBS primers generated 628 PCR amplicons, with an average of 209 amplicons. The average polymorphism level varied from 34% to 40% for all studied samples. Resolution analysis of the PBS primers showed all of them to have high or medium polymorphism levels, which varied from 0.763 to 0.965. Results of the molecular analysis of variance showed that the general biodiversity of A. ledebourianum populations is due to interpopulation (67%) and intrapopulation (33%) differences. The revealed genetic diversity was higher in the most distant population of A. ledebourianum LD64, located on the Sarymsakty ridge of Southern Altai. This is the first genetic diversity study of the endemic species A. ledebourianum using DNA profiling approaches. This work allowed us to collect new genetic data on the structure of A. ledebourianum populations in the Altai for subsequent development of preservation strategies to enhance the reproduction of this relict species. The results will be useful for the conservation and exploitation of this species, serving as the basis for further studies of its evolution and ecology.

Source Level Attribution: DNA Profiling from the ABAcard® HemaTrace® Kit

ABAcard® HemaTrace® kits have been used for crime scene stains for confirmation of human blood for many years. However, when the stain is too small to allow for separate testing, confirmatory testing may be forgone to preference DNA analysis. This can lead to court challenges as to the biological source and therefore probative value of the DNA profile. This research aimed to develop a protocol for DNA analysis of a minute blood stain subsequent to HemaTrace® testing. Stains were collected and subjected to HemaTrace® testing. Swabs were then removed from the HemaTrace® buffer solution and processed. DNA yields and STR DNA profiles were analysed for both quantity and quality. Full profiles were reliably obtained from stains with diameters of 0.6 mm–0.7 mm, reflecting DNA concentrations between 0.0036 ng/μL and 0.007 ng/μL, varying according to substrate characteristics. However, stains below a diameter of 0.6 mm should proceed directly for DNA profiling. This protocol was also successfully performed on blood stains which had undergone UV irradiation, although use of the reporting peak height threshold (lower than the routine analytical threshold) was required to obtain useable profiles. We have been able to demonstrate a protocol which, with minor adjustments to crime scene procedures, allows for both the confirmation of the presence of human blood, together with the generation of useful DNA profiles.