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American Journal of Human Genetics Apr 1992
Topics: DNA Fingerprinting; Homicide; Humans; Vermont
PubMed: 1550130
DOI: No ID Found -
Journal of Assisted Reproduction and... Nov 2014To validate a novel and more practical system for trophectoderm DNA fingerprinting which reliably distinguishes sibling embryos from each other.
PURPOSE
To validate a novel and more practical system for trophectoderm DNA fingerprinting which reliably distinguishes sibling embryos from each other.
METHODS
In this prospective and blinded study two-cell and 5-cell samples from commercially available sibling cell lines and excess DNA from trophectoderm biopsies of sibling human blastocysts were evaluated for accurate assignment of relationship using qPCR-based allelic discrimination from 40 single nucleotide polymorphisms (SNPs) with low allele frequency variation and high heterozygosity.
RESULTS
Cell samples with self relationships averaged 95.1 ± 5.9 % similarity. Sibling relationships averaged 57.2 ± 5.9 % similarity for all 40 SNPs, and 40.8 ± 8.2 % similarity for the 25 informative SNPs. Assignment of relationships was accomplished with 100 % accuracy for cell lines and embryos.
CONCLUSIONS
These data demonstrate the first trophectoderm qPCR-based DNA fingerprinting technology capable of unequivocal discrimination of sibling human embryos. This methodology will empower research and development of new markers of, and interventions that influence embryonic reproductive potential.
Topics: Blastocyst; Cell Line; DNA Fingerprinting; Gene Frequency; Heterozygote; Humans; Polymorphism, Single Nucleotide; Prospective Studies; Real-Time Polymerase Chain Reaction
PubMed: 25129376
DOI: 10.1007/s10815-014-0315-z -
International Journal of Molecular... Sep 2022Body fluid identification at crime scenes can be crucial in retrieving the appropriate evidence that leads to the perpetrator and, in some cases, the victim. For this...
Body fluid identification at crime scenes can be crucial in retrieving the appropriate evidence that leads to the perpetrator and, in some cases, the victim. For this purpose, immunochromatographic tests are simple, fast and suitable for crime scenes. The potential sample is retrieved with a swab, normally a cotton swab, moistened in a specific buffer. Nonetheless, there are other swab types available, which have been proven to be efficient for DNA isolation and analysis. The aim of this study is to evaluate the efficiency of different swab types for body fluid identification as well as DNA isolation and characterization. Fifty microliters of human saliva were deposited in three different types of fabric (denim, cotton, and polyester). After 24 h at room temperature, samples were recovered by applying three different swab types, and the tests were performed. Subsequently, total DNA was recovered from the sample buffer. Cotton swabs performed worse in denim and cotton fabrics in both immunochromatography tests and DNA yield. No differences were observed for polyester. In contrast, and except for two replicates, it was possible to obtain a full DNA profile per fabric and swab type, and to identify the mtDNA haplogroup. In this paper, the impact of swab types on body fluid identification through the application of immunochromatographic tests is analyzed for the first time. This work corroborates previous research related to the influence of swab types in nuclear DNA isolation and characterization.
Topics: DNA Fingerprinting; DNA, Mitochondrial; Humans; Polyesters; Saliva; Specimen Handling
PubMed: 36142599
DOI: 10.3390/ijms231810686 -
Legal Medicine (Tokyo, Japan) May 2024For human identification, the quality and quantity of DNA must be sufficient for amplification and analysis. When DNA extraction from bone tissues and teeth is required,...
For human identification, the quality and quantity of DNA must be sufficient for amplification and analysis. When DNA extraction from bone tissues and teeth is required, the optimal skeletal elements should be selected as samples for DNA extraction because DNA yield differs among elements. Recently, some studies have reported that a high quantity of high-quality DNA can be extracted from the small cancellous bones of the hands and feet. In this study, we evaluated the effectiveness of small cancellous bones in the human identification of skeletal remains in routine forensic genetic casework. Cancellous bones [phalanges, (meta)carpal bones, and (meta)tarsal bones)] and the cortical bones (femur and petrous bones) and teeth, which have generally been recommended as samples, were collected from the same individuals that needed identifying using DNA analysis in our laboratory. The quantity of DNA from small cancellous bones tended to be higher than that from cortical bones, and the quality from the former was as high as that from the latter. This study showed that in routine forensic casework, the small cancellous bones of the hands and feet should be actively selected as samples for DNA testing.
Topics: Humans; DNA; Forensic Genetics; Male; Bone and Bones; DNA Fingerprinting; Female; Cortical Bone; Middle Aged; Tooth; Adult; Aged; Forensic Anthropology; Cancellous Bone
PubMed: 38280273
DOI: 10.1016/j.legalmed.2024.102415 -
Forensic Science International. Genetics Jul 2022The association of body fluids/cell types and donors in mixed biological traces is an important, but challenging task required to evaluate the value of evidence given...
The association of body fluids/cell types and donors in mixed biological traces is an important, but challenging task required to evaluate the value of evidence given forensic propositions concerning the source of the DNA. The linking of a DNA profile with evidence from presumptive tests or RNA analysis is not straightforward. Coding region SNPs (cSNPs) are a novel type of evidential markers that are both cell type specific and individual specific. They thereby provide a direct link between a donor and a body fluid in mixed biological stains. In this proof-of-concept paper we consider the evaluation of cSNP profiles given source level propositions and explore the use of the open-source software EuroForMix to compute likelihood ratios. The discrimination power of the cSNPs for various body fluids is investigated with simulations. We provide case examples where the type of biological material is questioned and where cSNP profiles can be used to assign a donor to a body fluid, and discuss how the results can be reported in court.
Topics: Body Fluids; DNA; DNA Fingerprinting; Forensic Genetics; Humans; Polymorphism, Single Nucleotide
PubMed: 35381476
DOI: 10.1016/j.fsigen.2022.102685 -
Poultry Science Apr 1996An experiment was conducted to estimate genetic parameters in six experimental and five commercial primary breeding turkey lines using DNA fingerprinting. Eighteen...
An experiment was conducted to estimate genetic parameters in six experimental and five commercial primary breeding turkey lines using DNA fingerprinting. Eighteen individual DNA samples per line were digested with an HaeIII restriction enzyme and hybridized with Jeffreys' 33.6 probe. The DNA fingerprints were analyzed with computer programs to measure band sharing (BS) and band frequencies. Within lines, BS ranged from 0.39 to 0.62 and reflected the history of the experimental lines. Among lines, BS ranged from 0.21 to 0.33 with an average of 0.26. The BS among the experimental lines reflected known relationships. All lines were subdivided based on indices of population subdivision. About 26 hypervariable loci were estimated from band frequencies. Average heterozygosity and genetic variability estimated from band frequencies were significantly different among lines and displayed a result very similar to the BS among lines. Genetic distance indices among lines were also significantly different and reflected known relationships between the experimental lines. The experimental selected lines displayed lower genetic diversity than did the other lines. The parameters measuring genetic diversity within lines had higher correlation coefficients among them than did the parameters between lines. The computer program used in this study made DNA fingerprinting easier to use in population analysis.
Topics: Animals; Blotting, Southern; Breeding; Computer Simulation; DNA; DNA Fingerprinting; DNA Restriction Enzymes; Female; Gene Deletion; Gene Frequency; Genetic Variation; Heterozygote; Male; Models, Genetic; Polymorphism, Restriction Fragment Length; Turkeys
PubMed: 8786931
DOI: 10.3382/ps.0750439 -
Electrophoresis Apr 2017Next generation sequencing (NGS) is the emerging technology in forensic genomics laboratories. It offers higher resolution to address most problems of human...
Next generation sequencing (NGS) is the emerging technology in forensic genomics laboratories. It offers higher resolution to address most problems of human identification, greater efficiency and potential ability to interrogate very challenging forensic casework samples. In this study, a trial set of DNA samples was artificially degraded by progressive aqueous hydrolysis, and analyzed together with the corresponding unmodified DNA sample and control sample 2800 M, to test the performance and reliability of the ForenSeq DNA Signature Prep kit using the MiSeq Sequencer (Illumina). The results of replicate tests performed on the unmodified sample (1.0 ng) and on scalar dilutions (1.0, 0.5 and 0.1 ng) of the reference sample 2800 M showed the robustness and the reliability of the NGS approach even from sub-optimal amounts of high quality DNA. The degraded samples showed a very limited number of reads/sample, from 2.9-10.2 folds lower than the ones reported for the less concentrated 2800 M DNA dilution (0.1 ng). In addition, it was impossible to assign up to 78.2% of the genotypes in the degraded samples as the software identified the corresponding loci as "low coverage" (< 50x). Amplification artifacts such as allelic imbalances, allele drop outs and a single allele drop in were also scored in the degraded samples. However, the ForenSeq DNA Sequencing kit, on the Illumina MiSeq, was able to generate data which led to the correct typing of 5.1-44.8% and 10.9-58.7% of 58 of the STRs and 92 SNPs, respectively. In all trial samples, the SNP markers showed higher chances to be typed correctly compared to the STRs. This NGS approach showed very promising results in terms of ability to recover genetic information from heavily degraded DNA samples for which the conventional PCR/CE approach gave no results. The frequency of genetic mistyping was very low, reaching the value of 1.4% for only one of the degraded samples. However, these results suggest that further validation studies and a definition of interpretation criteria for NGS data are needed before implementation of this technique in forensic genetics.
Topics: DNA Fingerprinting; Electrophoresis, Capillary; Forensic Genetics; Genotype; Humans; Hydrolysis; Reagent Kits, Diagnostic; Reproducibility of Results; Sequence Analysis, DNA
PubMed: 28078776
DOI: 10.1002/elps.201600290 -
BioTechniques Sep 2022For microscopic investigation, archaeological bone samples are often embedded in Biodur epoxy resin. This study wants to test whether it is possible to extract DNA...
For microscopic investigation, archaeological bone samples are often embedded in Biodur epoxy resin. This study wants to test whether it is possible to extract DNA suitable for PCR amplification from this sample type. For eight individuals a set of samples - each consisting of a Biodur-embedded femur sample, a native femur sample and a control sample of different anatomical origin - were submitted to organic DNA extraction. The extraction success was tested by autosomal short tandem repeat amplification. Seven out of eight Biodur-embedded femur samples revealed successful amplification results. If Biodur-embedded bone material exists from earlier microscopic investigations, our results encourage the use of this sample type as a source for genetic research.
Topics: DNA; DNA Fingerprinting; DNA, Ancient; Epoxy Resins; Humans; Microsatellite Repeats
PubMed: 36066013
DOI: 10.2144/btn-2022-0056 -
Croatian Medical Journal Jun 2017A collaborative exercise with several institutes was organized by the Forensic DNA Service (FDNAS) and the Institute of the Legal Medicine, 2nd Faculty of Medicine,...
AIM
A collaborative exercise with several institutes was organized by the Forensic DNA Service (FDNAS) and the Institute of the Legal Medicine, 2nd Faculty of Medicine, Charles University in Prague, Czech Republic, with the aim to test performance of different laboratories carrying out DNA analysis of relatively old bone samples.
METHODS
Eighteen laboratories participating in the collaborative exercise were asked to perform DNA typing of two samples of bone powder. Two bone samples provided by the National Museum and the Institute of Archaelogy in Prague, Czech Republic, came from archeological excavations and were estimated to be approximately 150 and 400 years old. The methods of genetic characterization including autosomal, gonosomal, and mitochondrial markers was selected solely at the discretion of the participating laboratory.
RESULTS
Although the participating laboratories used different extraction and amplification strategies, concordant results were obtained from the relatively intact 150 years old bone sample. Typing was more problematic with the analysis of the 400 years old bone sample due to poorer quality.
CONCLUSION
The laboratories performing identification DNA analysis of bone and teeth samples should regularly test their ability to correctly perform DNA-based identification on bone samples containing degraded DNA and potential inhibitors and demonstrate that risk of contamination is minimized.
Topics: Bone and Bones; Czech Republic; DNA; DNA Fingerprinting; Forensic Genetics; Humans
PubMed: 28613037
DOI: 10.3325/cmj.2017.58.203 -
Forensic Science International. Genetics Mar 2022The identification of human remains belonging to missing persons is one of the main challenges for forensic genetics. Although other means of identification can be...
The identification of human remains belonging to missing persons is one of the main challenges for forensic genetics. Although other means of identification can be applied to missing person investigations, DNA is often extremely valuable to further support or refute potential associations. When reference DNA samples cannot be collected from personal items belonging to a missing person, a direct DNA identification cannot be carried out. However, identifications can be made indirectly using DNA from the missing person's relatives. The ranking of likelihood ratio (LR) values, which measure the fit of a missing person for any given pedigree, is often the first step in selecting candidates in a DNA database. Although implementing DNA kinship matching in a national environment is feasible, many challenges need to be resolved before applying this method to an international configuration. In this study, we present an innovative and intuitive method to perform international DNA kinship matching and facilitate the comparison of DNA profiles when the ancestry is unknown or unsure and/or when different marker sets are used. This straightforward method, which is based on calculations performed with the DNA matching software BONAPARTE, Worldwide allele frequencies and tailored cutoff logLR thresholds, allows for the classification of potential candidates according to the strength of the DNA evidence and the predicted proportion of adventitious matches. This is a powerful method for streamlining the decision-making process in missing person investigations and DVI processes, especially when there are low numbers of overlapping typed STRs. Intuitive interpretation tables and a decision tree will help strengthen international data comparison for the identification of reported missing individuals discovered outside their national borders.
Topics: DNA; DNA Fingerprinting; Databases, Nucleic Acid; Decision Making; Forensic Genetics; Gene Frequency; Humans; Likelihood Functions; Pedigree
PubMed: 34871915
DOI: 10.1016/j.fsigen.2021.102634