Rapid fractionation and isolation of whole blood components in samples obtained from a community-based setting

Amy Weckle, Allison E. Aiello, Monica Uddin, Sandro Galea, Rebecca M. Coulborn, Richelo Soliven, Helen Meier, Derek E. Wildman

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Article numbere52227
JournalJournal of Visualized Experiments
Volume2015
Issue number105
DOIs
StatePublished - Nov 30 2015

Keywords

  • Community-based study
  • DNA isolation
  • Issue 105
  • Medicine
  • Non-clinical setting
  • PBMC isolation
  • RNA isolation
  • Whole blood

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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