Influence of formalin fixation duration on RNA quality and quantity from formalin‐fixed paraffin‐embedded hepatocellular carcinoma tissues
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- Kenji Amemiya
- Genome Analysis Center Yamanashi Central Hospital Yamanashi Japan
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- Yosuke Hirotsu
- Genome Analysis Center Yamanashi Central Hospital Yamanashi Japan
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- Yuki Nagakubo
- Genome Analysis Center Yamanashi Central Hospital Yamanashi Japan
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- Hitoshi Mochizuki
- Genome Analysis Center Yamanashi Central Hospital Yamanashi Japan
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- Toshio Oyama
- Pathology Division, Laboratory Department Yamanashi Prefectural Central Hospital Yamanashi Japan
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- Masao Omata
- Department of Gastroenterology Yamanashi Central Hospital Yamanashi Japan
書誌事項
- 公開日
- 2023-11-07
- 資源種別
- journal article
- 権利情報
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- http://onlinelibrary.wiley.com/termsAndConditions#vor
- DOI
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- 10.1111/pin.13385
- 公開者
- Wiley
この論文をさがす
説明
<jats:title>Abstract</jats:title><jats:p>Analyzing RNA samples from formalin‐fixed paraffin‐embedded (FFPE) tissues is essential for precision medicine. We investigated RNA quantity and quality from FFPE tumor tissues fixed in formalin for various times and compared sequencing metrics from next‐generation sequencing (NGS). Hepatocellular carcinoma (HCC) tissues were fixed in 10% neutral buffered formalin (1–240 h) and FFPE blocks were prepared. Total RNA was extracted, and the quantity and quality were assessed using the NanoDrop, Qubit and Bioanalyzer. After preparing sequencing libraries, NGS was performed on the Oncomine Dx Multi‐CDx system. Total RNA yields of all samples met the threshold required for NGS, but longer fixation times resulted in decreased total RNA and long RNA fragment (>200 nt) yields. NGS analysis showed fewer sequencing reads of internal control genes from RNA with longer fixation times. RNA extracted from FFPE blocks stored for 500 days had reduced RNA yield and quality compared with RNA obtained from FFPE blocks prepared immediately. In conclusion, short and over‐fixation should be avoided because of their negative impact on sequencing quality. Fixation process should be finished promptly within recommended guidelines (6–72 h) for cancer patients.</jats:p>
収録刊行物
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- Pathology International
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Pathology International 73 (12), 593-600, 2023-11-07
Wiley
