Levels of 5′ RNA tags in plasma and buffy coat from EDTA blood increase with time

Fiona Marriage, Philip Day, F. Salway, P. J R Day, W. E R Ollier, T. C. Peakman

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Background: For biological sample banking it is important to precisely document sample treatment prior to extraction and storage. A major variable is the interval between blood sampling and subsequent processing and storage. We have determined the relationship between this time interval and frequency of 5′ transcript tags. This study was designed to establish guidelines for collecting RNA from blood in prospective studies and ensure maximum availability of RNA analytes. Methods: Venous blood was collected from 40 healthy volunteers. Samples were processed immediately, 12, 24 and 36 h post collection and buffy coat and/or plasma removed. Total RNA was extracted and reverse transcribed, assays were optimized and levels of 5′ RNA tags quantified by qPCR. Results: Stably expressed reference genes were selected to examine 5′ tags in plasma and buffy coat blood fractions. Whole blood was processed at various time points post collection to determine the affect on the presence and stability of 5′ RNA tags. A significant increase (P
    Original languageEnglish
    Pages (from-to)11-15
    Number of pages4
    JournalInternational Journal of Epidemiology
    Volume37
    Issue number1
    DOIs
    Publication statusPublished - 1 Apr 2008

    Keywords

    • Buffy coat
    • Gene expression
    • Plasma
    • Real-time PCR
    • RNA

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