The effects of storage temperature and duration of blood samples on DNA and RNA qualities

PLoS One. 2017 Sep 19;12(9):e0184692. doi: 10.1371/journal.pone.0184692. eCollection 2017.

Abstract

DNA and RNA samples from blood are the common examination target for non-invasive physical tests and/or biomedical studies. Since high-quality DNA and RNA samples guarantee the correctness of these tests and/or studies, we investigated the effects of storage temperature and storage duration of whole blood on DNA and RNA qualities. Subjects were enrolled to donate blood samples which were stored for different durations and at different temperatures, followed by the examinations on RNA quality, qPCR, DNA quality and DNA methylation. For RNA, we observed obvious quality decline with storage duration longer than 24 hours. Storage at low temperature does not keep RNA samples from degradation. And, storing whole blood samples in freezer dramatically damage RNA. For DNA, quality decline was not observed even with storage duration for 15 days. However, DNA methylation significantly altered with storage duration longer than three days. Storage duration within 24 hours is critical for collecting high-quality RNA samples for next-generation sequencing (NGS) assays (RIN≧8). If microarray assays are expected (RIN≧7), storage duration within 32 hours is acceptable. Although DNA is resistant within 15 days when kept in whole blood, DNA quantity dramatically decreases owing to WBC lysis. In addition, duration for more than three days significantly alter DNA methylation status, globally and locally. Our result provides a reference for dealing with blood samples.

MeSH terms

  • Blood Preservation*
  • DNA / isolation & purification
  • DNA / metabolism*
  • DNA Methylation
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Leukocytes / cytology
  • Leukocytes / metabolism
  • RNA / isolation & purification
  • RNA / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sequence Analysis, RNA
  • Temperature
  • Time Factors

Substances

  • RNA
  • DNA

Grants and funding

This article was funded by the grants from the Ministry of Science and Technology, Taiwan (MOST 106-2311-B-182A-002) and from Chang Gung Memorial Hospital (CMRPG8F0591 and CLRPG8D0113).