One of the most challenging points for ancient DNA studies on human remains is that analysing procedures involve partial or total destruction of the samples, which usually are unique and irreplaceable. This is what prevents museums, anthropologists or archaeologists from giving samples for genetic investigation. So that it is interesting to find an analysing method without destroying them. In this study, it was carried out the evaluation of a non-destructive extraction DNA protocol on 4 Neolithic individuals, from Can Gambús (Sabadell, Spain) and 4 pre Bell Beaker period individuals, from Los Cercados (Valladolid, Spain). Along this work, it was discussed the efficiency of the non-destructive protocol, valuing the obtained result, and the evidences state of preservation after the whole procedure.

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Kinship investigations such as paternity are currently solved using sets of (commercially available) highly polymorphic autosomal short tandem repeats (STRs), which lead to powerful likelihood ratios (LR). Still, some difficult cases arise whenever the kinship is much more remote or if the alternative hypotheses are not correctly formulated due to the lack of information (for e.g. there is an unknown relationship between the alleged and the true fathers). In these situations, beyond the routinely used marker set, laboratories usually enlarge the number and/or the type of markers analysed. Among these, autosomal indels and X-chromosome STRs have gained popularity. The aim of this study was to compare the results obtained after complementing an initial set of autosomal STRs with indels or with X-chromosome-specific STRs in simulated paternity cases where the alleged father is a close relative of the real one. Results show that in paternity cases where a low number of incompatibilities are observed, the best strategy is to increase the number of autosomal STRs under analysis. Nevertheless, if these are not available, our study globally shows that in father-daughter duos, a set of 12 X-STRs is more advantageous than 38 highly diverse autosomal biallelic markers. Additionally, the usefulness of X-STRs was also evaluated in cases where only a close relative of the alleged parent (father or mother) is available for testing. For those situations where these markers have the power to exclude, strong LR values are obtained. In the remaining cases, LRs are usually weak and sometimes the results are more likely under the wrong kinship hypothesis.

The remains in the LTB tomb were not a traditional nuclear family (father, mother and son/daughter) and it was probably a tomb where two women, one of them pregnant, were buried.

Through this study we compared two DNA extraction methods for skeletal or dental human samples. For such\ud task there were analysed 38 samples from 19 individuals, selecting two samples from each one. When it was\ud possible, we selected dental complete samples, without cracks or cavities, and also with a natural light colour.\ud When there were no dental samples available, there were selected preferably skull or diaphysis of long bone.\ud The two samples from each individual were processed individually in two different laboratories (Laboratories\ud 1 and 2). There were employed a different DNA extraction methodology in each laboratory, applying in\ud Laboratory 1 the protocol proposed by Rohland and Hofreiter (2009), and in Laboratory 2 a commercial kit for\ud purification. Finally, in order to compare the efficiency of both methodologies, in Laboratory 2, aDNA quantification\ud by Real Time PCR (RTPCR) was performed, by the amplification of two different size mitochondrial DNA\ud fragments. In this way, it was possible to evaluate the efficiency of each protocol, and to discuss advantages and\ud disadvantages of each one

The present study focuses on the genetic analysis of skeletal human remains exhumed from a ritual burial located\ud in Los Cercados Chalcolithic site (3970 ± 60 BP) (Valladolid, Central Spain). In this burial different pottery and\ud animal remains were found, configuring a complex ritual, accompanied by scarce human remains, concretely a\ud maxilla and three skulls without maxilla.\ud The most striking aspect of these human remains was the different impact trauma signs on the back side of the\ud skulls. The anthropological analysis established that the skulls were typical feminine, The bad state of preservation\ud of the maxilla did not allowed to assign this to any of the three skulls. So, it was not possible to\ud determine the number of individuals by anthropological methodology.\ud However, we could determine the number of individuals by the genetic analysis of autosomal STRs and\ud mitochondrial DNA on the skeletal remains. It was possible to assign the maxilla to one of other three human\ud skulls. On the other hand, we have been able to verify the sex of each individual by molecular analysis. Finally, a\ud kinship analysis among the individuals was performed using a specific software (Familias 3.0), resulting in a\ud possible sibling relationship between two of the individuals

One of the most effective ways for human identification is the genetic analysis of the crime scene biological evidence(s). However, such studies could be expensive, and it could take too long before the genetic profile is finally obtained (1).In the present work, we have made modifications to the conventional DNA extraction protocol (1), in order to perform a "direct" amplification, trying to obtain the optimal conditions for generating a fast and reliable genetic profile. In order to do so, we employed different fresh blood and saliva samples, and carried out an extraction with the Prep-n-Go™ buffer (ThermoFisher™Scientific, Foster City, USA) during 30 s of incubation. Then the extract was collected to perform the autosomal amplification with GlobalFiler™ Express PCR Amplification Kit (ThermoFisher™Scientific, Foster City, USA). The quality of the new procedure always took into account the minimum standards of quality, estimated by the allele's height in RFUs. On the other hand, the aptitude of our newly proposed protocol to keep the required quality, performing sensibility, reproducibility and contamination tests was evaluated.We were able to conclude that it was possible to obtain a reliable genetic profile performing a "direct" amplification, although the best amplification conditions varied according to the type of sample. In general, the best obtained results were determined in a range of 1 μl extract volume, and it was possible to obtain a genetic profile in 90 min.

Although biological relationships are a universal reality for all human beings, the concepts of “family” and “family bond” depend on both the geographic region and the historical moment to which they refer. However, the concept of “family” can be determinant in a large variety of societies, since it can influence the lines of succession, inheritances and social relationships, as well as where and with whom an individual is buried. The relation between a deceased person and other members of a community, other individuals of the same necropolis, or even with those who are buried in the same tomb can be analysed from the genetic point of view, considering different perspectives: archaeological, historical, and forensic. In the present work, the concepts of “family” and “kinship” are discussed, explaining the relevance of genetic analysis, such as nuclear and lineage markers, and their contribution to genealogical research, for example in the heritage of surnames and Y-chromosome, as well as those cases where some discrepancies with historical record are detected, such as cases of adoption. Finally, we explain how genetic genealogical analyses can help to solve some cold cases, through the analysis of biologically related relatives.