A novel multiplex assay for simultaneously analysing 13 rapidly mutating Y-STRs.

A multiplex polymerase chain reaction (PCR) assay (RM-Yplex) was developed which is capable of simultaneously amplifying 13 recently introduced rapidly mutating Y-STR markers (RM Y-STRs). This multiplex assay is expected to aid human identity testing in forensic and other applications to improve differentiating unrelated males and allow separating related males. The 13 RM Y-STR markers included in the multiplex are: DYF387S1, DYF399S1, DYF403S1ab, DYF404S1, DYS449, DYS518, DYS526ab, DYS547, DYS570, DYS576, DYS612, DYS626 and DYS627. This study reflects the proof of concept to analyse all currently known RM Y-STRs simultaneously and describes the optimization of the multiplex assay. The RM-Yplex assay generated complete RM Y-STR profiles down to 62.5pg of male template DNA, and from male-female DNA mixtures at all ratios tested. We herewith introduce and make available for widespread use in forensic and anthropological studies, an effective and sensitive single multiplex assay for simultaneous genotyping of 13 RM Y-STRs.

Development of a multiplex system to assess DNA persistence in taphonomic studies.

In this study, we have developed a PCR multiplex that can be used to assess DNA degradation and at the same time monitor for inhibition: primers have been designed to amplify human, pig, and rabbit DNA, allowing pig and rabbit to be used as experimental models for taphonomic research, but also enabling studies on human DNA persistence in forensic evidence. Internal amplified controls have been added to monitor for inhibition, allowing the effects of degradation and inhibition to be differentiated. Sequence data for single-copy nuclear recombination activation gene (RAG-1) from human, pig, and rabbit were aligned to identify conserved regions and primers were designed that targeted amplicons of 70, 194, 305, and 384 bp. Robust amplification in all three species was possible using as little as 0.3 ng of template DNA. These have been combined with primers that will amplify a bacterial DNA template within the PCR. The multiplex has been evaluated in a series of experiments to gain more knowledge of DNA persistence in soft tissues, which can be important when assessing what material to collect following events such as mass disasters or conflict, when muscle or bone material can be used to aid with the identification of human remains. The experiments used pigs as a model species. When whole pig bodies were exposed to the environment in Northwest England, DNA in muscle tissue persisted for over 24 days in the summer and over 77 days in the winter, with full profiles generated from these samples. In addition to time, accumulated degree days (ADD) were also used as a measure that combines both time and temperature-24 days was in summer equivalent to 295 ADD whereas 77 days in winter was equivalent to 494 ADD.

Demographic characteristics of elite Ethiopian endurance runners.

INTRODUCTION: The dominance of East-African athletes in distance running remains largely unexplained; proposed reasons include favorable genetic endowment and optimal environmental conditions. PURPOSE: To compare the demographics of elite Ethiopian athletes with the general Ethiopian population and assess the validity of reports linking running long distances to school with endurance success. METHODS: Questionnaires, administered to 114 members (male and female) of the Ethiopian national athletics team and 111 Ethiopian control subjects (C) obtained information on place of birth, language, distance and method of travel to school. Athletes were separated into three groups according to athletic discipline: marathon (M; N = 34); 5,000-10,000 m (5-10 km; N = 42); and other track and field athletes (TF; N = 38). Frequency differences between groups were assessed using contingency chi-square tests. RESULTS: Regional distributions of marathon athletes differed from controls (P < 0.001) and track and field athletes (P = 0.013), but not the 5- to 10-km athletes (P = 0.21). The 5- to 10-km athletes also differed from controls (P < 0.001). Marathon athletes exhibited excess from the regions of Arsi and Shewa (M: 73%; 5-10 km: 43%; TF: 29%; C: 15%). The language distribution of marathon athletes differed from all groups (P < 0.001), with a predominance of languages of Cushitic origin (M: 75%, 5-10 km: 52%, TF: 46%, C: 30%). A higher proportion of marathon athletes ran to school (M: 68%; 5-10 km: 31%; TF: 16%; C: 24%) and traveled greater distances. CONCLUSION: Elite endurance athletes are of a distinct environmental background in terms of geographical distribution, ethnicity, and also having generally traveled farther to school, often by running. These findings may reflect both environmental and genetic influences on athletic success in Ethiopian endurance athletes.