Optimized protocol for the extraction of RNA and DNA from frozen whole blood sample stored in a single EDTA tube
Abstract 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 fo...
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Nature Portfolio
2021
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oai:doaj.org-article:212124c659ab4ed4bcf4e9068db01cdf2021-12-02T18:53:08ZOptimized protocol for the extraction of RNA and DNA from frozen whole blood sample stored in a single EDTA tube10.1038/s41598-021-96567-22045-2322https://doaj.org/article/212124c659ab4ed4bcf4e9068db01cdf2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-96567-2https://doaj.org/toc/2045-2322Abstract 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.Hirotaka YamagataAyumi KobayashiRyouichi TsunedomiTomoe SekiMasaaki KobayashiKosuke HagiwaraChong ChenShusaku UchidaGo OkadaManabu FuchikamiToshiharu KamishikiryoJun-ichi IgaShusuke NumataMakoto KinoshitaTakahiro A. KatoRyota HashimotoHiroaki NaganoYasumasa OkamotoShuichi UenoTetsuro OhmoriShin NakagawaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021) |
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Medicine R Science Q Hirotaka Yamagata Ayumi Kobayashi Ryouichi Tsunedomi Tomoe Seki Masaaki Kobayashi Kosuke Hagiwara Chong Chen Shusaku Uchida Go Okada Manabu Fuchikami Toshiharu Kamishikiryo Jun-ichi Iga Shusuke Numata Makoto Kinoshita Takahiro A. Kato Ryota Hashimoto Hiroaki Nagano Yasumasa Okamoto Shuichi Ueno Tetsuro Ohmori Shin Nakagawa Optimized protocol for the extraction of RNA and DNA from frozen whole blood sample stored in a single EDTA tube |
| description |
Abstract 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. |
| format |
article |
| author |
Hirotaka Yamagata Ayumi Kobayashi Ryouichi Tsunedomi Tomoe Seki Masaaki Kobayashi Kosuke Hagiwara Chong Chen Shusaku Uchida Go Okada Manabu Fuchikami Toshiharu Kamishikiryo Jun-ichi Iga Shusuke Numata Makoto Kinoshita Takahiro A. Kato Ryota Hashimoto Hiroaki Nagano Yasumasa Okamoto Shuichi Ueno Tetsuro Ohmori Shin Nakagawa |
| author_facet |
Hirotaka Yamagata Ayumi Kobayashi Ryouichi Tsunedomi Tomoe Seki Masaaki Kobayashi Kosuke Hagiwara Chong Chen Shusaku Uchida Go Okada Manabu Fuchikami Toshiharu Kamishikiryo Jun-ichi Iga Shusuke Numata Makoto Kinoshita Takahiro A. Kato Ryota Hashimoto Hiroaki Nagano Yasumasa Okamoto Shuichi Ueno Tetsuro Ohmori Shin Nakagawa |
| author_sort |
Hirotaka Yamagata |
| title |
Optimized protocol for the extraction of RNA and DNA from frozen whole blood sample stored in a single EDTA tube |
| title_short |
Optimized protocol for the extraction of RNA and DNA from frozen whole blood sample stored in a single EDTA tube |
| title_full |
Optimized protocol for the extraction of RNA and DNA from frozen whole blood sample stored in a single EDTA tube |
| title_fullStr |
Optimized protocol for the extraction of RNA and DNA from frozen whole blood sample stored in a single EDTA tube |
| title_full_unstemmed |
Optimized protocol for the extraction of RNA and DNA from frozen whole blood sample stored in a single EDTA tube |
| title_sort |
optimized protocol for the extraction of rna and dna from frozen whole blood sample stored in a single edta tube |
| publisher |
Nature Portfolio |
| publishDate |
2021 |
| url |
https://doaj.org/article/212124c659ab4ed4bcf4e9068db01cdf |
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