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Chromenes and quinolines are recognized as important scaffolds in medicinal chemistry. Herein, the efficient use of chromene- and quinoline-3-carbaldehydes to synthesize other valuable heterocycles is described. These carbaldehydes are obtained in excellent yields through the Vilsmeyer-Haack reaction of flavanones and azaflavanones. Protocols towards the synthesis of new heterocycles, such as 3H-chromeno[3&ndash, c]quinolines, (Z/E)-2-aryl-4-chloro-3-styryl-2H-chromenes, and (E)-2-aryl-4-chloro-3-styrylquinoline-1(2H)-carbaldehydes were established. Altogether, we demonstrate the value of chromene- and quinoline-3-carbaldehydes as building blocks.

A collaborative European DNA Profiling (EDNAP) Group exercise was undertaken to assess the performance of an earlier described SNaPshot™-based screening assay (denoted mini-mtSNaPshot) (Weiler et al., 2016) [1] that targets 18 single nucleotide polymorphism (SNP) positions in the mitochondrial (mt) DNA control region and allows for discrimination of major European mtDNA haplogroups. Besides the organising laboratory, 14 forensic genetics laboratories were involved in the analysis of 13 samples, which were centrally prepared and thoroughly tested prior to shipment. The samples had a variable complexity and comprised straightforward single-source samples, samples with dropout or altered peak sizing, a point heteroplasmy and two-component mixtures resulting in one to five bi-allelic calls. The overall success rate in obtaining useful results was high (97.6%) given that some of the participating laboratories had no previous experience with the typing technology and/or mtDNA analysis. The majority of the participants proceeded to haplotype inference to assess the feasibility of assigning a haplogroup and checking phylogenetic consistency when only 18 SNPs are typed. To mimic casework procedures, the participants compared the SNP typing data of all 13 samples to a set of eight mtDNA reference profiles that were described according to standard nomenclature (Parson et al., 2014) [2], and indicated whether these references matched each sample or not. Incorrect scorings were obtained for 2% of the comparisons and derived from a subset of the participants, indicating a need for training and guidelines regarding mini-mtSNaPshot data interpretation.

This paper describes a paternity case investigation that became a maternity investigation, in which the mother of two dizygotic male twins referred having received a bone marrow transplant, donated by her sister, seven years before, due to a sickle cell disease. The study of 17 STRs with different commercial kits revealed two maternal inconsistencies with one child and one inconsistency with the other. The alleged father was excluded as the biological father without genetic information of the mother. After receiving our report, the court decided to investigate the maternity relative to the children. Supplementary studies with more STRs and with other mother biological tissues revealed distinct genetic profiles in blood, epithelial cells and hair roots. The profile obtained from the hair roots was consistent with being the mother of her children. This case shows the importance of accurate information about the medical condition of all the involved persons, along with the collection of hair roots, especiall...

In forensic genetic laboratories, low template DNA (LTDNA) samples can be a challenge in the attempting to obtain genetic profiles with the most information possible. In this study, volume reduction on amplification reaction was tested in the kit Globalfiler™, with the intend to optimize the results in LTDNA samples. To carry out this study, several samples extracted with PrepFiler ® were diluted successively to very low concentration values. All samples and respective dilutions were amplified with Globalfiler™, for both reaction volumes of 12,5μL and 25μL. Capillary electrophoresis was performed on the ABI PRISM ® 3500 Genetic Analyzer and the HID files generated were analysed using GeneMapper ® ID-X v.1.4. Forensic DNA samples with very low values of DNA concentration were also tested in the same conditions. The results indicate that the reduction of volume in amplification reaction with Globalfiler™ can often produce higher quality profiles, resulting as an alternative to enhance allelic recovery.

The Treaty of Prum signed on 27 May 2005, by seven Member States of European Union (EU), sought to improve cross-border security cooperation in combating terrorism, cross-border crime and illegal immigration. For this purpose, provisions were set for automated exchanges of data regarding DNA, fingerprints and vehicle registration plates. Later, other Member States expressed their intention to accede to the Prum Convention, including Portugal. Portuguese forensic DNA profiles database was implemented in 2010, although only in 2015 Portugal started the automated DNA data exchange with other countries. From that time until now, it has connected to a large number of countries in order to fulfil the purpose for which it was intended when accessed the Prum Convention, following the guidelines of EU Decision 2008/615/JHA. It is currently exchanging DNA data automatically with 21 Member States. The objective of this work is to describe the evolution of the Portuguese forensic DNA database under Prum Treaty over the past few years, analysing several information such as the number and type of genetic profiles shared, the number of hits that occurred with other countries, as well as other relevant information.

Massively Parallel Sequencing (MPS) allows to sequence the total mitochondrial DNA (mtDNA) through specific panels. Human Mitochondria Panel is one of the QIAseq Targeted DNA panels developed by QIAGEN for MPS, which have an optimized chemistry based on the integration of unique molecular indices (UMIs). However, the major problem is the analysis of the MPS data and, therefore, it became necessary to create bioinformatics tools that would enable mtDNA analysis such as the AQME tool to be used with the QIAGEN Biomedical Genomics Workbench software. In this study, a coverage comparison workflow was tested to verify the coverage differences between two mtDNA panels, QIAseq Human Mitochondria Panel and Precision ID mtDNA Whole Genome Panel. The preliminary results showed a significant decrease in the Precision ID coverage, after the bioinformatics removal of PCR artifacts. For the QIAseq panel, this coverage variation was lower, since the UMIs enable to know more accurately the initial number of molecules in the original mtDNA sequence.

Improvements in scientific techniques allowed the analysis of limited amounts of DNA and the testing of not only body fluids but also touch DNA. The possibility of DNA transfer often raises the question of how the DNA was deposited: directly or by indirect transfer? Therefore, it is important to comprehend the factors that can influence it. Our study shows that background DNA and DNA indirect transfer are in fact a reality, having the potential to interfere with target DNA detections, and corroborates that area of contact and type of material are of importance on DNA transfer.

After a validation study developed in the Forensics Genetic and Biology Service of Coimbra, Portugal, we were able to demonstrate the ability of the GlobalFiler ® PCR Amplification kit in direct amplification of reference samples with consistent and reproducible results. Since this kit is optimized for casework samples, we decided to test it on forensic samples, in particular on fabrics stains. The selected samples had already been processed according to the methodologies normally used for this type of samples included automated extraction, quantification and amplification, followed by capillary electrophoresis, resulting in complete profiles. To develop our study, samples were collected from regions of the fabrics that had previously shown/proven to have biological evidence. The chosen samples were directly amplified for the GlobalFiler ® PCR Amplification kit, followed by capillary electrophoresis on the ABI PRISM ® 3500 Genetic Analyzer. The results obtained with this methodology revealed full and well balanced genetic profiles. This study intends to show that in some cases this approach can be an alternative, when we are in the presence of biological stains on fabrics, that allows to obtain consistent results in a short period of time.

The GlobalFiler ® PCR Amplification system uses 6-dye fluorescent chemistry to enable multiplexing of 21 autosomal STRs, 1 Y-STR, 1 Y-indel and the sex-determining marker amelogenin. Validation studies were conducted to assess the performance of the GlobalFiler ® casework kit for typing reference samples through a direct amplification. The findings support that the GlobalFiler ® PCR Amplification Kit produces reliable results from blood and buccal reference DNA samples.

Degradation can be identified due to the presence of a downward slope across the electropherogram, where larger loci have a lower probability to be amplified and thus a major chance of dropping out. The introduction of the degradation index (DI) of Quantifiler ® Trio and the optimization of the available amplification kits have been improving the genotyping success of these samples. Casework samples were amplified with GlobalFiler™ and PowerPlex ® Fusion 6C, according to the quantification results. Quantification and amplification results demonstrated that it is important to consider DI and DNA concentration values to define downstream procedure and also that the genetic information provided by both amplification kits could be useful for degraded samples profiling.

Genetic profiles obtained from gynaecological swabs or a victim's underwear usually are imbalanced mixtures of female vs male profile. Performing differential DNA extraction prior to DNA amplification may result in an enriched sperm fraction and a consequently isolated male profile. The aim of this work was to test the ability of Sampletype I-sep DL extraction system to separate epithelial from sperm cells, followed by purification with Prepfiler. Although the I-sep did not always provide an isolated male profile, in almost half of the casework samples resulted in a profile with the male contribution in majority.

Mitochondrial DNA (mtDNA) has some unique features that make it a useful forensic marker such as absence of recombination across many generations, maternal inheritance, accumulation of mutations over time and multiple copies per cell. The application of massively parallel sequencing (MPS) to forensic genetics has expanded in recent years due to its potential to dramatically increase sample throughput, workflow efficiency and detection resolution. Recently, Applied Biosystems™ developed the Precision ID mtDNA Whole Genome Panel, specifically for mtDNA forensic applications. The preliminary results showed that mtDNA haplotypes obtained with MPS were consistent with those previously obtained with traditional Sanger sequencing, regardless of the extraction method used and pool concentration of libraries.

This study analyzes the allelic frequency distribution of 17 STRs contained in the AmpF l STR Identifiler (Applied Biosystems) and PowerPlex16 System (Promega) commercial kits for two large population samples from the Azores archipelago (Portugal) ( N =475) and from Central Portugal ( N =2125). Likewise, it includes a comparative study among the population groups analyzed in this paper and those which history points out as originating from the first settlers of the Azores. All loci were highly polymorphic. The Central Portuguese area and the Azores archipelago population samples are in Hardy–Weinberg equilibrium for the 17 markers analyzed.

Background: The non-recombining portion of the Y chromosome represents a valuable tool for the study of human population history. The haploid, non-crossing-over behaviour of Y chromosome gives potentials in many aspects resembling the maternally inherited mtDNA. The advantages for forensic casework are obvious. The haploid state makes mixture patterns much simpler; in female/male stain mixtures, pure male DNA profiles may be obtained. Y-chromosome markers are also of extraordinary value to solve selected paternity cases. Methods: Y-chromosomal biallelic markers have been investigated in a population sample from central Portugal (n=117). By combining the allelic state of 10 biallelic markers (YAP-DYS287, SRY-8299, 92R7, 12f2, SRY-1532, SRY-2627, Tat, SY81, M9, LLY22g), we could define the haplogroup to which each sample belonged. Results: This population defined 10 different haplogroups, haplogroup 1 being the most frequent. Conclusion: This study showed a substantial similarity with other Iberian studies; however, this population presents a larger diversity. http://www.sciencedirect.com/science/article/B7581-47W664D-1J4/1/ec7f5cd37a8d6f852a3e3bad356295b6

The aim of the present work was to study the origin of paternal and maternal lineages in Guinea-Bissau population, inferred by phylogeographic analyses of mtDNA and Y chromosome defined haplogroups. To determine the male lineages present in Guinea-Bissau, 33 unrelated males were typed using a PCR-SNaPshot multiplex based method including 24 Y-SNPs, which characterize the main haplogroups in sub-Saharan Africa and Western Europe. In the same samples, 17 Y-STRs (included in the YFiler kit, Applied Biosystems) were additionally typed. The most frequent lineages observed were E1b1a (xE1b1a4,7)-M2 (68%) and E1a-M33 (15%). The European haplogroup R1b1-P25 was represented with a frequency of 12%. The two hypervariable mtDNA regions were sequenced in 79 unrelated individuals from Guinea-Bissau, and haplogroups were classified based on control region motifs using mtDNA manager. A high diversity of haplogroups was determined in our sample being the most frequent haplogroups characteristic of populations from sub-Saharan Africa, namely L2a1 (15%), L3d (13%), L2c (9%), L3e4 (9%), L0a1 (8%), L1b (6%) and L1c1 (6%). None of the typical European haplogroups (H, J and T) were found in the present sample of Guinea-Bissau. From our results, it is possible to confirm that Guinea-Bissau presents a typically West African profile, marked by a high frequency of the Y chromosome haplogroup E1b1a(xE1b1a4,7)-M2 and a high proportion of mtDNA lineages belonging to the sub-Saharan specific sub-clusters L1 to L3 (89%). A small European influx has been also detected, although restricted to the male lineages.

A part of a decomposed human body from an individual was found at his home, together with the decomposed bodies of his 3 dogs. The disappearance of half of the human body was hardly explained. Hypothetically the dogs, starving, could have eaten their owner after his death. All the bodies were recovered for autopsy. Small bone fragments were recovered in one dog's stomach and identified as human by anthropological analysis. DNA was extracted and cytochrome b gene analysis was made in order to determine their origin, confirmed as human. Genetic identification allowed achieving an eight mini-STR profile with MiniFiler (Applied Biosystems) identical to the bone material collected at the victim's autopsy, confirmed that the dogs had effectively eaten their owner. The results showed that it is possible to obtain nuclear DNA in samples subjected to gastric acids and the combination of different techniques allowed us to determine, in each step, the most convenient workflow for the remains identification.

The AmpFlSTR ® MiniFiler™ PCR amplification kit (Applied Biosystems), a new available 8-miniSTR and the sex determining marker Amelogenin multiplex, includes the most common problematic loci (above 200bp) of the AmpFlSTR ® Identifiler™ PCR amplification kit: FGA, D21S11, D18S51, D13S317, D7S820, D16S539, CSF1PO and D2S1338. Several casework samples with different DNA contents were tested. Results allowed to complete partial Identifiler™ profiles and additional information was achieved in low copy number (LCN) samples, revealing that this miniSTR kit can improve identification of compromised samples.

Haplotypes of 111 individuals from La Paz have been studied by the analysis of the polymorphisms in mtDNA control region, HV1 and HV2 regions, in order to intend the genetic diversity. The aim was to created a population database, assess genetic interpopulation variability and classify haplogroups. We had identify 99 haplotypes with 125 polymorphic nucleotide positions, 93 haplotypes are unique. Nucleotide and sequence diversity are estimated in 0.016068±0.008186 and 0.9988, respectively. Haplogroup distribution is as follows: 47.75% B; 14.41% C; 6.31% A; 4.5% D; 6.31% A, C, or D; 23.42% undetermined or compound haplogroups. Rate of heteroplasmy is 37.8% in HV1 and 53.2% in HV2.

The new quantification kit known as Quantifiler ® Trio is introduced as more robust and sensitive compared to the Quantifiler ® Duo, providing additional information in the analysis of forensic samples. Validation studies were designed to evaluate the sensitivity of the Quantifiler ® Trio kit and its performance with mixture and degraded samples.

We examine the flow in a horizontal Hele-Shaw cell in which the undisturbed unidirectional flow at infinity is required to stream around a vertical cylinder spanning the gap between the two (horizontal) plates of the cell. A combination of matched asymptotic expansions and numerical methods is employed to elucidate the structure of the boundary layer near the surface of the cylinder. The two length scales of the problem are the gap, h, and the length of the body, l; it is assumed that h/l

The aim of this study was the analysis of the origin of maternal lineage (mitochondrial DNA) and paternal lineage (Y Short Tandem Repeats) of Azores Islands population comparing our data with other populations from Europe and Africa. The comparison of maternal and paternal lineages from Azores Islands with other lineages from Europe and Africa was performed using the Arlequin software version 2.000.

Biological traces from dogs and other domestic animals can be found in a crime scene related to suspects or victims being themselves the protagonists of the forensic case. According to Portuguese Legislation, the individual identification of dogs dangerous or potentially dangerous is obligatory. It seems important to follow this subject with the implementation of genetic identification. So, this work has the aim to study STRs of DNA for individual genetic identification of dogs. DNA extracted from saliva of different dogs breed was amplified with StockMarks® Kit—Canine Genotyping.

We performed a study of the allelic frequency distribution of 17 STRs along each of the islands of Azores archipelago to look out for statistical differences. From the global samples, we selected those individuals whose both parents were born in the same island. There were no significant differences among islands exception to Flores with some P-values not reaching 0.05.

Crime scene evidence and samples from decomposed or skeletal remains have usually low copy number (LCN), with templates containing less than 100 pg of DNA. The choice of the best extraction methods as well as sensitive and robust STRs is crucial to obtain good results. However, in many situations, it is also necessary to make adaptations to standard protocols, such as increase in the number of amplification cycles (even when commercial kits are used) or the injection time in capillary sequencers. A set of four amplifications was made, with 28 (two of them), 32 and 34 cycles. To the samples with 28 PCR amplification cycles was added more enzyme and two other amplifications with 4 and 6 cycles was performed. Results were analysed and discussed.

The nonlinear interaction between a pair of symmetric, oblique, and spatial instability modes is studied in the long-wave limit using asymptotic methods. The base flow is taken to be a supersonic mixing layer whose Mach number is such that the corresponding vortex sheet is marginally stable according to Miles' criterion. It is shown that the amplitude of the mode obeys a nonlinear integro-differential equation. Numerical solutions of this equation show that, when the obliqueness angle is less than pi/4, the effect of the nonlinearity is to enhance the growth rate of the instability. The solution terminates in a singularity at a finite streamwise location. This result is reminiscent of that obtained in the vicinity of the neutral point by other authors in several different types of flows. On the other hand, when the obliqueness angle is more than pi/4, the streamwise development of the amplitude is characterized by a series of modulations. This arises from the fact that the nonlinear term in the amplitude equation may be either stabilizing or destabilizing, depending on the value of the streamwise coordinate. However, even in this case the amplitude of the disturbance increases, though not as rapidly as in the case for which the angle is less than pi/4. Quite generally then, the nonlinear interaction between two oblique modes in a supersonic mixing layer enhances the growth of the disturbance.

The aim of this study was to analyse the distribution of Y-chromosomal haplotypes and haplogroups found in central Portugal. In this work, we analysed 102 unrelated individuals of central Portugal. Combining the allelic state of 10 biallelic markers (YAP, SRY-8299, 92R7, 12f2, SRY-1532, SRY-2627, Tat, SY81, M9, LLY22g), we defined the haplogroup to which each sample belonged. Y-chromosomal haplotypes were defined by 16 Y-Short Tandem Repeats (STR) (DYS19, DYS3855 a/b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS460, DYS461, GATA A10, GATA C4 and GATA H4). This population study defined 8 different haplogroups and 101 different haplotypes, where 100 haplotypes were unique and 1 was found in two apparently unrelated individuals, both belonging to the same haplogroup.

Our casuistic in paternity testing showed several isolated incompatibilities present in different STRs like FES, SE33, vWA, among others, include homozygote and heterozygote situations, and appear both in father–child and mother–child pairs. As we assume, in paternity testing, that the mother is the real one, most of incompatibility situations are assigned to a mutation in putative father. In fact, only sequencing allows the real explanation for the event.

Using the assumptions of an incompressible and viscous flow at large Reynolds number, we derive the evolution equation for the wave action density of an instability wave travelling on top of a laminar free-shear flow. The instability is considered to be viscous; the purpose of the present work is to include the cumulative effect of the (locally) small viscous correction to the wave, over length and time scales on which the underlying base flow appears inhomogeneous owing to its viscous diffusion. As such, we generalize our previous work for inviscid waves. This generalization appears as an additional (but usually non-negligible) term in the equation for the wave action. The basic structure of the equation remains unaltered.

We consider analytically the evolution of a three-dimensional wave packet generated by an impulsive source in a mixing layer. The base flow is assumed to be spreading due to viscous diffusion. The analysis is restricted to small disturbances (linearized theory). A suitable high-frequency ansatz is used to describe the packet; the key ele­ments of this description are a complex phase and a wave action density. It is found that the product of this density and an infinitesimal material volume convecting at the local group velocity is not conserved: there is a cohtinuous interaction be­tween the base flow and the wave action. This interaction is determined by suitable mode-weighted averages of the second and fourth derivatives of the base-flow veloc­ity profile. Although there is some tendency for the dominant wavenumber in the packet to shift from the most unstable value towards the neutral value, this shift is quite moderate. In practice, wave packets do not become locally neutral in a diverg­ing base flow (as do instability modes), therefore, they are expected to grow more suddenly than pure instability modes and do not develop critical layers. The group velocity is complex; the full significance of this is realized by analytically continuing the equations for the phase and wave action into a complex domain. The implications of this analytic continuation are discussed vis-a-vis the secondary instabilities of the packet: very small-scale perturbations on the phase can grow very rapidly initially, but saturate later because most of the energy in these perturbations is convected away by the group velocity. This remark, as well as the one regarding critical layers, has consequences for nonlinear theories.

We study the flow generated by a small circular cylinder in a mixing layer. The cylinder is executing an oscillatory translation whose frequency is within the range of unstable frequencies of the shear layer. The smallness of the cylinder is measured by the ratio of its radius to the characteristic thickness of the layer. This (small) ratio serves as the expansion parameter for our theory; the flow naturally divides into inner and outer regions. The former is in the immediate vicinity of the cylinder and the latter is the far field which contains the instability waves. The solution to this problem is obtained by the method of matched asymptotic expansion. One objective is to study the dependence of this solution on various parameters such as the frequency of oscillation, velocity ratio, etc., and thus shed light on the associatedreceptivity. Other objectives deal with a restatement of causality and with the hydrodynamic field near the streamwise location of the cylinder. We find that receptivity is a strong function of frequency and velocity ratio and that the local hydrodynamic field may be quite large. Causality is restated in terms of the well-known exponential integral.

Mitochondrial DNA analysis is very useful for the interpretation of the history of human migration and to estimate the frequency of a haplotype in the forensic context. From a human settlement perspective, La Paz area is greatly interesting since the first planned city of the region is located there. Samples from 110 individuals from La Paz were studied analysing the polymorphisms in the D-loop, hypervariable region I (HVI) and hypervariable region II (HVII) in order to verify the genetic diversity. The aim of this study was to start the creation of a population database in order to obtain the genetic interpopulation variability and classify haplotypes into characteristic haplogroups of South America. A total of 97 different haplotypes were identified, 90 being unique, expressed by 122 polymorphic nucleotide positions. Nucleotide and sequence diversity were estimated to be 0.015 ± 0.0075 and 0.996, respectively. Haplogroup distribution in the samples was 57.27% B4, 19.09% C1, 10.00% A2, 3.64% D1, 2.73% D4h3, 1.82% H, and 0.91% for each of the haplogroups A4, B4c1a, CZ, D4J, M7a and M8/N9b. The rate of length heteroplasmy was 36.36% in HVI and 52.73% in HVII. Phylogenetic analysis reveals proximity to the Korean, Chilean aboriginal, Japanese and Australian populations. The estimated genetic variability of the studied population was high, suggesting an early settlement.

17 Y-chromosome STR loci (DYS19, DYS389I, DYS389II, DYS390, DYS456, DYS391, DYS392, DYS393, DYS385 a/b, DYS458, DYS439, DYS635, GATA H4.1, DYS437, DYS438 and DYS448) were determined for 303 unrelated males, living in Central Portugal, using the AmpFlSTR ® YFiler™ PCR Amplification kit (Applied Biosystems). A total of 287 different haplotypes were found, 272 being unique. The overall haplotype diversity (HD) was determined as 0.9996, a value similar to other YFiler data sets. Y-STR polymorphisms in Central Portugal population, using YFiler, provide a powerful discrimination tool for routine forensic applications.

The stability of a family of tanh mixing layers is studied at large Mach numbers using perturbation methods. It is found that the eigenfunction develops a multilayered structure, and the eigenvalue is obtained by solving a simplified version of the Rayleigh equation (with homogeneous boundary conditions) in one of these layers which lies in either of the external streams. Our analysis leads to a simple hypersonic similarity law which explains how spatial and temporal phase speeds and growth rates scale with Mach number and temperature ratio. Comparisons are made with numerical results, and it is found that this similarity law provides a good qualitative guide for the behaviour of the instability at high Mach numbers.In addition to this asymptotic theory, some fully numerical results are also presented (with no limitation on the Mach number) in order to explain the origin of the hypersonic modes (through mode splitting) and to discuss the role of oblique modes over a very wide range of Mach number and temperature ratio.

In 2003, an aircraft accident occurred in the Madeira island north coast, in which 10 persons (4 female and 6 male) have died. STRs (autosomic and Y-chromosome) and mtDNA were made in order to identify the recovered human body remains and results compared with relatives.

This work aimed to study the rapidly mutating Y-STRs in a population of Portugal Centre. The results showed that these markers are highly polymorphic, observing that each sample had a unique haplotype.

A B S T R A C T Challenging biological samples found in crime scenes are often brought to our lab. Several factors, such as degradation and the presence of inhibitors, can difficult the analysis of these samples. Chelating resin, silica membranes, silica-coated magnetic beads and paramagnetic resin were DNA extraction techniques used in this study. Our aim was to find out the DNA extraction method more suitable to overcome problems raised by these samples and to enable the identification of its genetic profile.

The AmpFlSTR ® Identifiler ® Direct PCR Amplification Kit is a short tandem repeat (STR) multiplex assay, optimized to amplify 16 loci directly from biological material (blood or buccal cell samples) stored in FTA paper, without resorting to DNA extraction. However, preliminary experiences have shown the versatility and robustness of this amplification kit, when applied to other biological sample supports. The aim of this research work is to analyze the results of this kit when applied to non-FTA paper or other support for biological samples such as a buccal swab.

AmpFlSTR ® NGM™ is a 16-locus multiplex kit claiming improved robustness than earlier generation AppliedBiosystems kits; NGM-SElect™ was released recently as an equivalent to NGM loci plus SE33 so it was determined if NGM-SElect can be an asset compared to NGM in low-level DNA samples.

The aim of the present work was to study the origin of paternal lineages in Angola population, namely in the three main ethnic groups, inferred by phylogeographic analyses of Y chromosome defined haplogroups. To determine the male lineages present in the Angola population studied, 112 unrelated males were typed using a PCR multiplex – minisequencing SnaPshot based method including 29Y-SNPs, which characterize the majority haplogroups in sub-Saharan Africa populations.

Mitochondrial DNA (mtDNA) has enormous potential in population genetics allowing the correlation and identification of the origin of individuals in a population. In forensic genetics mtDNA enables the identification of the matrilineal lineage from degraded samples. Genetic testing can be performed using mtDNA noncoding region and/or mtDNA coding region. However, to apply this inclusive approach as evidence in legal practice is necessary to determine the genetic structure of populations. Traditional mtDNA sequencing and Single Nucleotide Polymorphisms (SNP) analysis techniques were combined to establish the mtDNA variability of the Azorean population and classify the haplotypes into haplogroups. Forty-eight haplotypes were identified. All mtDNA haplotypes were included into specific haplogroups: forty-three haplotypes belonging to macrohaplogroup R; three haplotypes belonging to macrohaplogroup N; and two haplotypes belonging to macrohaplogroup M.

Samples analysed in this study suffered from afflictions, such as low level DNA, degradation and PCR inhibitors: samples with induced degraded conditions and real crime scene samples, from a wide range of crimes. Results obtained with NGM SElect™ PCR amplification kit and PowerPlex ® ESI 17 system were compared, to verify the behaviour of both kits in the presence of challenging samples.

Samples of 70 individuals from 3 ethnic groups from Angola (Bakongos, Kimbundu and Ovimbundu) were studied analysing the polymorphisms of the entire mitochondrial control region in order to verify the genetic diversity. A total of 68 different haplotypes were identified, 66 being unique. Sequence diversity was estimated to be 1.0000±0.0113 and nucleotide diversity for Bakongo, Kimbundu and Ovimbundu ethnic groups were 0.015556±0.007970, 0.016772±0.008517 and 0.016020±0.008411, respectively. It was possible to include the majority of the mtDNA haplotypes into a specific African mtDNA haplogroup.

TM A B S T R A C T A variety of challenging biological samples, including blood stains, saliva, semen, hair, bones, finger nails, among others, are often a part of our casework investigation. In this study, semen, blood samples and saliva swabs were extracted by several methods in order to optimize and validate the Prepfiler Express TM Extraction kit and the AutoMate Express DNA Extraction System. Results obtained with the robot (using silica-coated magnetic beads) were compared with methods based on a chelating resin, silica membranes and paramagnetic resin.