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.

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.

Summary This paper examines the identification of kinship relations in archaeological multiple burials and advocates the application of different methods and lines of research to clarify such issues in relation to funerary practices. Recognizing family relationships – an important task in research on prehistoric societies – is especially complicated and interpretations have often been made without an adequate empirical basis. Bioarchaeological, isotopic and DNA analyses applied to the triple burial of Los Tolmos (Cogotas I archaeological culture, Iberian Bronze Age) have provided direct information on this issue. In this respect, the new results also imply the need to consider gender constructs in greater depth and to be more open‐minded towards other forms of relationship in the past beyond the traditional heteronormative nuclear family.

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

Con motivo de presentar dos tumbas individuales en fosa inéditas, se aborda un estudio de conjunto sobre las prácticas funerarias del Calcolítico Precampaniforme en la submeseta norte española. Pese a su carácter mayoritario en el registro arqueológico, se discute la representatividad como norma de este tipo de sepulturas; se rastrean comportamientos propios de rituales de enterramiento en dos tiempos, preludio de los documentados en este mismo espacio en la Edad del Bronce; se aporta información sobre el ADN mitocondrial y sexo molecular de uno de los difuntos, y se pone el énfasis, por vez primera en el Calcolítico de la cuenca del Duero, en el protagonismo de los perros en el mundo funerario. Por último, la proximidad de las dos nuevas sepulturas a sendos dólmenes –Los Zumacales (Valladolid) y La Velilla (Palencia)? da pie a debatir sobre el inicio de la ‘postvida’ megalítica en el valle medio del Duero.

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.

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