Non-Governmental Organization Archaia (http://www.archaia.cz) carried out the rescue archaeological research at Knezeves near Prague in 1998. Most of dating objects in Knezeves come from the period of Late and Final Bronze Age. The approximately 3,000 years old set, which included 11 human remains from three settlement features, was collected for the study. First, gender was determined according to anthropological characteristics. Ancient DNA from bones was extracted by the phenol-chloroform procedure and N-phenacetylthiazolum bromide reagent. Polymerase chain reaction amplification of AMEL XY, part of amelogenin gene, with subsequent polyacrylamide gel electrophoresis and Short Tandem Repeats analysis followed. DNA profiles of skeletal remains were obtained by the fragmentation analysis of autosomal short tandem repeat markers. Genetic profiles showed us whether individuals from Knezeves were in mutual relationship (parent - descendant). The congruence of results in sex determination supported reliability of genetic methods, which are suitable for sex determination of fragmental and subadult skeletal remains.

• MeSH
• archeologie metody   MeSH
• dějiny starověku MeSH
• DNA analýza   MeSH
• kosti a kostní tkáň chemie   MeSH
• lidé MeSH
• mikrosatelitní repetice MeSH
• polymerázová řetězová reakce metody   MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA metody   MeSH
• Check Tag
• dějiny starověku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• historické články MeSH
• odvolaná publikace MeSH

Research on ancient and forensic DNA is related in many ways, and the two fields must deal with similar obstacles. Therefore, communication between these two communities has the potential to improve results in both research fields. Here, we present the insights gained in the ancient DNA community with regard to analyzing DNA from aged skeletal material and the potential use of the developed protocols in forensic work. We discuss the various steps, from choosing samples for DNA extraction to deciding between classical PCR amplification and massively parallel sequencing approaches. Based on the progress made in ancient DNA analyses combined with the requirements of forensic work, we suggest that there is substantial potential for incorporating ancient DNA approaches into forensic protocols, a process that has already begun to a considerable extent. However, taking full advantage of the experiences gained from ancient DNA work will require comparative studies by the forensic DNA community to tailor the methods developed for ancient samples to the specific needs of forensic studies and case work. If successful, in our view, the benefits for both communities would be considerable.

• MeSH
• DNA fingerprinting MeSH
• DNA * genetika   MeSH
• lidé MeSH
• nekrotická degradace DNA MeSH
• senioři MeSH
• soudní genetika MeSH
• starobylá DNA * MeSH
• Check Tag
• lidé MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Dna je velice staré onemocnění, které lidstvo pronásleduje už několik tisíc let. První popisy interpretované jako projevy dny se objevují již v egyptských lékařských papyrech ze 3. tisíciletí před Kristem. V antickém světě se příčinami, diagnostikou a léčením dny zabývala celá řada lékařů, mezi nimi např. Hippokratés z Kósu, Dioklés z Karystu nebo Claudios Galénos. Dna v personifikované podobě (jako bohyně Podagra) si však našla místo i v antické mytologii a kultuře. Ze starověku se také dochovalo několik kosterních ostatků osob, které trpěly dnou.

Gout is a very old disease, which exists for thousands of years. The first descriptions interpreted as the symptoms of gout can be found already in the Egyptian medical papyri dating to the 3rd mill. BC. In the Ancient world, many physicians dealt with the causes, diagnostics and the treatments of gout, such as Hippocrates of Cos, Diocles of Carystus or Claudios Galenos. A personified gout (as the goddess Podagra) is also to be found in the Ancient mythology and culture. Several human remnants of the people suffering from gout are preserved from the Antiquity as well.

• MeSH
• dějiny lékařství MeSH
• dějiny starověku * MeSH
• dna (nemoc) * dějiny   MeSH
• lidé MeSH
• Check Tag
• dějiny starověku * MeSH
• lidé MeSH
• Publikační typ
• historické články MeSH

Working with mitochondrial DNA from highly degraded samples is challenging. We present a whole mitogenome Illumina-based sequencing method suitable for highly degraded samples. The method makes use of double-stranded library preparation with hybridization-based target enrichment. The aim of the study was to implement a new user-friendly method for analysing many ancient DNA samples at low cost. The method combines the Swift 2S™ Turbo library preparation kit and xGen® panel for mitogenome enrichment. Swift allows to use low input of aDNA and own adapters and primers, handles inhibitors well, and has only two purification steps. xGen is straightforward to use and is able to leverage already pooled libraries. Given the ancient DNA is more challenging to work with, the protocol was developed with several improvements, especially multiplying DNA input in case of low concentration DNA extractions followed by AMPure® beads size selection and real-time pre-capture PCR monitoring in order to avoid cycle-optimization step. Nine out of eleven analysed samples successfully retrieved mitogenomes. Hence, our method provides an effective analysis of whole mtDNA, and has proven to be fast, cost-effective, straightforward, with utilisation in population-wide research of burial sites.

• MeSH
• analýza nákladů a výnosů MeSH
• genom mitochondriální * MeSH
• lidé MeSH
• mitochondriální DNA genetika   MeSH
• polymerázová řetězová reakce MeSH
• soudní genetika metody   MeSH
• starobylá DNA * MeSH
• vysoce účinné nukleotidové sekvenování metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

AIM: To develop novel DNA extraction and typing procedure for DNA identification of the 7th century human remains, determine the familiar relationship between the individuals, estimate the Y-chromosome haplogroup, and compare the Y-chromosome haplotype with the contemporary populations. METHODS: DNA from preserved femur samples was extracted using the modified silica-based extraction technique. Polymerase chain reaction amplification was performed using human identification kits MiniFiler, Identifiler, and Y-filer and also laboratory-developed and validated Y-chromosome short tandem repeat (STR) pentaplexes with short amplicons. RESULTS: For 244A, 244B, 244C samples, full autosomal DNA profiles (15 STR markers and Amelogenin) and for 244D, 244E, 244F samples, MiniFiler profiles were produced. Y-chromosome haplotypes consisting of up to 24 STR markers were determined and used to predict the Y-chromosome haplogroups and compare the resulting haplotypes with the current population. Samples 244A, 244B, 244C, and 244D belong to Y-chromosome haplogroup R1b and the samples 244E and 244F to haplogroup G2a. Comparison of ancient haplotypes with the current population yielded numerous close matches with genetic distance below 2. CONCLUSION: Application of forensic genetics in archaeology enables retrieving new types of information and helps in data interpretation. The number of successfully typed autosomal and Y-STR loci from ancient specimens in this study is one of the largest published so far for aged samples.

• MeSH
• dějiny starověku MeSH
• DNA fingerprinting metody   MeSH
• genetické markery MeSH
• genotyp MeSH
• geny vázané na chromozom Y MeSH
• lidé MeSH
• mikrosatelitní repetice MeSH
• polymerázová řetězová reakce MeSH
• soudní antropologie MeSH
• soudní genetika MeSH
• zkameněliny MeSH
• Check Tag
• dějiny starověku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• historické články MeSH
• práce podpořená grantem MeSH

AIMS: Ancient DNA (aDNA) extracted from historical bones is damaged and fragmented into short segments, present in low quantity, and usually copurified with microbial DNA. A wide range of DNA quantification methods are available. The aim of this study was to compare the five most common DNA quantification methods for aDNA. MATERIALS AND METHODS: Quantification methods were tested on DNA extracted from skeletal material originating from an early medieval burial site. The tested methods included ultraviolet (UV) absorbance, real-time quantitative polymerase chain reaction (qPCR) based on SYBR(®) green detection, real-time qPCR based on a forensic kit, quantification via fluorescent dyes bonded to DNA, and fragmentary analysis. Differences between groups were tested using a paired t-test. RESULTS: Methods that measure total DNA present in the sample (NanoDrop(™) UV spectrophotometer and Qubit(®) fluorometer) showed the highest concentrations. Methods based on real-time qPCR underestimated the quantity of aDNA. The most accurate method of aDNA quantification was fragmentary analysis, which also allows DNA quantification of the desired length and is not affected by PCR inhibitors. CONCLUSIONS: Methods based on the quantification of the total amount of DNA in samples are unsuitable for ancient samples as they overestimate the amount of DNA presumably due to the presence of microbial DNA. Real-time qPCR methods give undervalued results due to DNA damage and the presence of PCR inhibitors. DNA quantification methods based on fragment analysis show not only the quantity of DNA but also fragment length.

• MeSH
• DNA genetika   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• lidé MeSH
• polymerázová řetězová reakce metody   MeSH
• starobylá DNA analýza   izolace a purifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Ancient DNA (aDNA) of Encephalitozoon intestinalis (Microsporidia, Fungi) was detected in archaeological material originated from New Town of Prague (Czech Republic) with the use of molecular methods. Microsporidial aDNA was found in 3 samples originating from 2 objects, in a well/cesspit (samples from layers from the 18th century) and in a well from the 18th/19th century. The ability to use molecular methods to detect microsporidia extends the range of paleoparasitological inquiry, and could contribute to a better understanding of parasites shared between human and animals.

• MeSH
• dějiny 18. století MeSH
• dějiny 19. století MeSH
• DNA fungální chemie   dějiny   izolace a purifikace   MeSH
• Encephalitozoon genetika   izolace a purifikace   MeSH
• encephalitozoonóza dějiny   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• sekvence nukleotidů MeSH
• zvířata MeSH
• Check Tag
• dějiny 18. století MeSH
• dějiny 19. století MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• historické články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

The Asiatic wild dog (Cuon alpinus), restricted today largely to South and Southeast Asia, was widespread throughout Eurasia and even reached North America during the Pleistocene. Like many other species, it suffered from a huge range loss towards the end of the Pleistocene and went extinct in most of its former distribution. The fossil record of the dhole is scattered and the identification of fossils can be complicated by an overlap in size and a high morphological similarity between dholes and other canid species. We generated almost complete mitochondrial genomes for six putative dhole fossils from Europe. By using three lines of evidence, i.e., the number of reads mapping to various canid mitochondrial genomes, the evaluation and quantification of the mapping evenness along the reference genomes and phylogenetic analysis, we were able to identify two out of six samples as dhole, whereas four samples represent wolf fossils. This highlights the contribution genetic data can make when trying to identify the species affiliation of fossil specimens. The ancient dhole sequences are highly divergent when compared to modern dhole sequences, but the scarcity of dhole data for comparison impedes a more extensive analysis.

• MeSH
• Canidae anatomie a histologie   klasifikace   genetika   MeSH
• fylogeneze * MeSH
• genom mitochondriální MeSH
• hybridizace genetická MeSH
• migrace zvířat MeSH
• mitochondriální DNA MeSH
• starobylá DNA * MeSH
• zkameněliny MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH

Ancient DNA from historical and subfossil wood has a great potential to provide new insights into the history of tree populations. However, its extraction and analysis have not become routine, mainly because contamination of the wood with modern plant material can complicate the verification of genetic information. Here, we used sapwood tissue from 22 subfossil pines that were growing c. 13 000 yr bp in Zurich, Switzerland. We developed and evaluated protocols to eliminate surface contamination, and we tested ancient DNA authenticity based on plastid DNA metabarcoding and the assessment of post-mortem DNA damage. A novel approach using laser irradiation coupled with bleaching and surface removal was most efficient in eliminating contaminating DNA. DNA metabarcoding confirmed which ancient DNA samples repeatedly amplified pine DNA and were free of exogenous plant taxa. Pine DNA sequences of these samples showed a high degree of cytosine to thymine mismatches, typical of post-mortem damage. Stringent decontamination of wood surfaces combined with DNA metabarcoding and assessment of post-mortem DNA damage allowed us to authenticate ancient DNA retrieved from the oldest Late Glacial pine forest. These techniques can be applied to any subfossil wood and are likely to improve the accessibility of relict wood for genome-scale ancient DNA studies.

• MeSH
• borovice klasifikace   genetika   MeSH
• dekontaminace MeSH
• DNA rostlinná genetika   izolace a purifikace   MeSH
• dřevo genetika   MeSH
• druhová specificita MeSH
• lesy * MeSH
• smrk genetika   MeSH
• zkameněliny * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Important gaps remain in our understanding of the spread of farming into Europe, due partly to apparent contradictions between studies of contemporary genetic variation and ancient DNA. It seems clear that farming was introduced into central, northern, and eastern Europe from the south by pioneer colonization. It is often argued that these dispersals originated in the Near East, where the potential source genetic pool resembles that of the early European farmers, but clear ancient DNA evidence from Mediterranean Europe is lacking, and there are suggestions that Mediterranean Europe may have resembled the Near East more than the rest of Europe in the Mesolithic. Here, we test this proposal by dating mitogenome founder lineages from the Near East in different regions of Europe. We find that whereas the lineages date mainly to the Neolithic in central Europe and Iberia, they largely date to the Late Glacial period in central/eastern Mediterranean Europe. This supports a scenario in which the genetic pool of Mediterranean Europe was partly a result of Late Glacial expansions from a Near Eastern refuge, and that this formed an important source pool for subsequent Neolithic expansions into the rest of Europe.

• MeSH
• běloši MeSH
• efekt zakladatele MeSH
• etnicita MeSH
• genetická variace * MeSH
• genom lidský * MeSH
• haplotypy MeSH
• lidé MeSH
• mitochondriální DNA analýza   MeSH
• starobylá DNA analýza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH
• Střední východ MeSH
• Středomoří MeSH