title: Rapid Fractionation and Isolation of Whole Blood Components in Samples Obtained from a Community-based Setting.
abstract: 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.
authors.affiliation: 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; aweckle@illinois.edu.
authors.forename: Amy
authors.initials: A
authors.lastname: Weckle
authors.affiliation: Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina.
authors.forename: Allison E
authors.initials: AE
authors.lastname: Aiello
authors.affiliation: Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign; Department of Psychology, University of Illinois at Urbana-Champaign.
authors.forename: Monica
authors.initials: M
authors.lastname: Uddin
authors.affiliation: Department of Epidemiology, Mailman School of Public Health, Columbia University.
authors.forename: Sandro
authors.initials: S
authors.lastname: Galea
authors.affiliation: Department of Epidemiology, University of Michigan School of Public Health.
authors.forename: Rebecca M
authors.initials: RM
authors.lastname: Coulborn
authors.affiliation: Center for Molecular Medicine and Genetics, Wayne State University School of Medicine.
authors.forename: Richelo
authors.initials: R
authors.lastname: Soliven
authors.affiliation: Department of Epidemiology, University of Michigan School of Public Health.
authors.forename: Helen
authors.initials: H
authors.lastname: Meier
authors.affiliation: 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.
authors.forename: Derek E
authors.initials: DE
authors.lastname: Wildman