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

doi:10.3791/52227

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Medicine

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JoVE Journal Medicine

Rapid Fractionation and Isolation of Whole Blood Components in Samples Obtained from a Community-based Setting

快速分馏和全血的成分的分离样品中从社区为基础的设置获得

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11:31 min

November 30, 2015

DOI: 10.3791/52227-v

Amy Weckle^1,2, Allison E. Aiello³, Monica Uddin^1,4, Sandro Galea⁵, Rebecca M. Coulborn⁶, Richelo Soliven⁷, Helen Meier⁶, Derek E. Wildman^1,2

¹Carl R. Woese Institute for Genomic Biology,University of Illinois at Urbana-Champaign, ²Department of Molecular and Integrative Physiology,University of Illinois at Urbana-Champaign, ³Department of Epidemiology, Gillings School of Global Public Health,University of North Carolina, ⁴Department of Psychology,University of Illinois at Urbana-Champaign, ⁵Department of Epidemiology, Mailman School of Public Health,Columbia University, ⁶Department of Epidemiology,University of Michigan School of Public Health, ⁷Center for Molecular Medicine and Genetics,Wayne State University School of Medicine

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我们概述了一种处理全血以获得各种成分以供进一步分析的方法。我们优化了一种简化的方案，能够在非临床环境中快速、高通量同时处理全血样本。

该程序的总体目标是提供一种简化的方案，实现快速的眼通量，同时处理全血样品。在非临床环境中，真空采血管处理以重叠方式进行，以有效分离 D-N-A-R-N-A 外周血单核细胞或 PBMC 血清的高质量组分以及血浆和红细胞的残留组分。处理从血清真空采血器的离心开始，从一个含有真空采血器的 fial 中取出等分试样，用于 fial 管的 DNA 分离离心。

然后完成 DNA 分离，在 DNA 分离后，可以从 K 两个 EDTA 真空中分离出白细胞衍生的 RNA。来自每个真空采血器的血液通过相应的过滤器并用 PBS 洗涤，然后用 RNA 稳定剂处理，返回到含有吸血器的 fial 中。吸出残留的血浆，并用 PBS 洗涤单核层。

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