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ID 117721
Yamagata, Hirotaka Yamaguchi University|Japan Science and Technology Agency
Kobayashi, Ayumi Yamaguchi University
Tsunedomi, Ryouichi Yamaguchi University
Seki, Tomoe Yamaguchi University|Japan Science and Technology Agency
Kobayashi, Masaaki Yamaguchi University
Hagiwara, Kosuke Yamaguchi University
Chen, Chong Yamaguchi University
Uchida, Shusaku Japan Science and Technology Agency|Kyoto University
Okada, Go Hiroshima University
Fuchikami, Manabu Hiroshima University
Kamishikiryo, Toshiharu Hiroshima University
Kato, Takahiro A. Kyushu University
Hashimoto, Ryota National Center of Neurology and Psychiatry
Nagano, Hiroaki Yamaguchi University
Okamoto, Yasumasa Hiroshima University
Ueno Ehime University
Nakagawa, Shin Yamaguchi University
Content Type
Journal Article
Cryopreservation of whole blood is useful for DNA collection, and clinical and basic research. Blood samples in ethylenediaminetetraacetic acid disodium salt (EDTA) tubes stored at − 80 °C are suitable for DNA extraction, but not for high-quality RNA extraction. Herein, a new methodology for high-quality RNA extraction from human blood samples is described. Quickly thawing frozen whole blood on aluminum blocks at room temperature could minimize RNA degradation, and improve RNA yield and quality compared with thawing the samples in a 37 °C water bath. Furthermore, the use of the NucleoSpin RNA kit increased RNA yield by fivefold compared with the PAXgene Blood RNA Kit. Thawing blood samples on aluminum blocks significantly increased the DNA yield by ~ 20% compared with thawing in a 37 °C water bath or on ice. Moreover, by thawing on aluminum blocks and using the NucleoSpin RNA and QIAamp DNA Blood kits, the extraction of RNA and DNA of sufficient quality and quantity was achieved from frozen EDTA whole blood samples that were stored for up to 8.5 years. Thus, extracting RNA from frozen whole blood in EDTA tubes after long-term storage is feasible. These findings may help advance gene expression analysis, as well as biomarker research for various diseases.
Journal Title
Scientific Reports
Springer Nature
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Medical Sciences
University Hospital