Weckle Amy, Aiello Allison E, Uddin Monica, Galea Sandro, Coulborn Rebecca M, Soliven Richelo, Meier Helen, Wildman Derek E
Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign; Department of Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign;
Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina.
J Vis Exp. 2015 Nov 30(105):52227. doi: 10.3791/52227.
Collection and processing of whole blood samples in a non-clinical setting offers a unique opportunity to evaluate community-dwelling individuals both with and without preexisting conditions. Rapid processing of these samples is essential to avoid degradation of key cellular components. Included here are methods for simultaneous peripheral blood mononuclear cell (PBMC), DNA, RNA and serum isolation from a single blood draw performed in the homes of consenting participants across a metropolitan area, with processing initiated within 2 hr of collection. We have used these techniques to process over 1,600 blood specimens yielding consistent, high quality material, which has subsequently been used in successful DNA methylation, genotyping, gene expression and flow cytometry analyses. Some of the methods employed are standard; however, when combined in the described manner, they enable efficient processing of samples from participants of population- and/or community-based studies who would not normally be evaluated in a clinical setting. Therefore, this protocol has the potential to obtain samples (and subsequently data) that are more representative of the general population.
在非临床环境中采集和处理全血样本,为评估有或无既往病史的社区居民提供了独特的机会。对这些样本进行快速处理对于避免关键细胞成分的降解至关重要。这里介绍的方法包括在大都市地区同意参与的参与者家中,通过一次采血同时分离外周血单核细胞(PBMC)、DNA、RNA和血清,并在采集后2小时内开始处理。我们已使用这些技术处理了1600多个血样,获得了一致的高质量材料,这些材料随后成功用于DNA甲基化、基因分型、基因表达和流式细胞术分析。所采用的一些方法是标准方法;然而,当以所述方式组合时,它们能够有效地处理来自基于人群和/或社区研究的参与者的样本,而这些参与者通常不会在临床环境中接受评估。因此,该方案有潜力获得更能代表一般人群的样本(以及随后的数据)。