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Quality assessment of RNA in long-term storage: The All Our Families biorepository

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  • Nikki L Stephenson
  • Kylie K Hornaday
  • Chelsea T A Doktorchik
  • Andrew W Lyon
  • Suzanne C Tough
  • Donna M Slater

Abstract

Background: The All Our Families (AOF) cohort study is a longitudinal population-based study which collected biological samples from 1948 pregnant women between May 2008 and December 2010. As the quality of samples can decline over time, the objective of the current study was to assess the association between storage time and RNA (ribonucleic acid) yield and purity, and confirm the quality of these samples after 7–10 years in long-term storage. Methods: Maternal whole blood samples were previously collected by trained phlebotomists and stored in four separate PAXgene Blood RNA Tubes (PreAnalytiX) between 2008 and 2011. RNA was isolated in 2011 and 2018 using PAXgene Blood RNA Kits (PreAnalytiX) as per the manufacturer’s instruction. RNA purity (260/280), as well as RNA yield, were measured using a Nanodrop. The RNA integrity number (RIN) was also assessed from 5–25 and 111–130 months of storage using RNA 6000 Nano Kit and Agilent 2100 BioAnalyzer. Descriptive statistics, paired t-test, and response feature analysis using linear regression were used to assess the association between various predictor variables and quality of the RNA isolated. Results: Overall, RNA purity and yield of the samples did not decline over time. RNA purity of samples isolated in 2011 (2.08, 95% CI: 2.08–2.09) were statistically lower (p

Suggested Citation

  • Nikki L Stephenson & Kylie K Hornaday & Chelsea T A Doktorchik & Andrew W Lyon & Suzanne C Tough & Donna M Slater, 2020. "Quality assessment of RNA in long-term storage: The All Our Families biorepository," PLOS ONE, Public Library of Science, vol. 15(12), pages 1-11, December.
    • Handle: RePEc:plo:pone00:0242404
    DOI: 10.1371/journal.pone.0242404
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