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

• Main Authors: Pei-Hsien Lin, 林配奾
• Other Authors: Chang-Yi Wu
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/3qg53u

碩士 === 國立中山大學 === 生物科學系研究所 === 106 === DNA and RNA quality are critical for biomedical studies such as microarray and NGS. 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 DNA and RNA qualities from blood. Blood samples 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 base on bioanalyzer and qPCR results. Storage at low temperature does not keep RNA samples from degradation. Storage of 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 status significantly altered with storage duration longer than three days through dot blot and pyromark. In summary, 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 white blood cells (WBCs) 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.