Paxgene blood rna kit handbook

For the current study, we selected 87 unrelated nonagenarians, 337 offspring and 321 partners belonging to 281 nuclear families (Table 1). These samples were randomly selected, but in such a way that age and gender were balanced between the groups and the age range was as large as possible. Only individuals without outlying cell counts (>3 SD below or above the standard error of the mean) were included. This subpopulation is representative for the whole LLS regarding disease prevalence and parameters involved in metabolic syndrome [9 (link), 41 (link)]. From these non-fasted individuals, peripheral blood was harvested using PAXgeneTM tubes (Qiagen, Venlo, The Netherlands). The tubes were frozen and kept at −20 °C for ~3–5 years. After thawing at room temperature for at least 2 h, total RNA was extracted from the approximately 2.5 ml of peripheral blood in each tube following the manufacturer’s recommended protocol (PAXgene Blood RNA Kit Handbook, Qiagen, Venlo, The Netherlands). The quality and integrity of the total RNA was evaluated on the 2100 Bioanalyzer (Agilent Technologies, Amstelveen, The Netherlands) and the concentration was measured using a NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). Quality criteria included a 28S/18S ratio, as measured by the 2100 Bioanalyzer, of at least 1.2, and a total RNA yield of at least 3 μg.