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学科主题: 基础医学
题名:
Deparaffinization and Lysis by Hydrothermal Pressure (Pressure Cooking) Coupled With Chaotropic Salt Column Purification: A Rapid and Efficient Method of DNA Extraction From Formalin-fixed Paraffin-embedded Tissue
作者: Zhong, Haohao1; Liu, Yan1; Talmor, Monica2; Wu, Bingquan1; Hui, Pei2
关键词: hydrothermal pressure ; DNA extraction ; FFPE
刊名: DIAGNOSTIC MOLECULAR PATHOLOGY
发表日期: 2013-03-01
DOI: 10.1097/PDM.0b013e318263f092
卷: 22, 期:1, 页:52-58
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Pathology
研究领域[WOS]: Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Pathology
关键词[WOS]: POLYMERASE-CHAIN-REACTION ; ARCHIVAL TISSUE ; PCR ; AMPLIFICATION ; RETRIEVAL ; SECTIONS
英文摘要:

We report a hydrothermal pressure method (pressure cooking) for simultaneous deparaffinization and lysis of formalin-fixed paraffin-embedded tissue followed by conventional chaotropic salt column purification to obtain high-quality DNA. Using this method, the release of DNA occurred within the first minute of treatment, reaching the maximum at 5 minutes. An optimal treatment window was between 5 and 30 minutes. The extracted DNA was of high quality as determined by the 260/280 absorbance ratios, and the quantity of DNA extracted was linear with the input tissue amount. In paired sample experiments (N=19), the quantity of DNA extracted by hydrothermal pressure treatment was comparable to that obtained through the conventional xylene deparaffinization and protease K digestion method. The integrity of the recovered DNA was also comparable, evidenced by polymerase chain reaction amplifications of variable-sized amplicons in tissue samples archived from 0.2 to 22 years (N=14). The high quality of DNA was further confirmed by polymerase chain reaction amplification and Sanger sequencing analysis of representative exons of the EGFR gene in human non-small cell lung cancer tissue samples. In summary, this novel method offers DNA release from formalin-fixed paraffin-embedded tissue with unprecedented simplicity, speed, biohazard safety, and cost-efficiency. Combined with chaotropic salt column purification, high-quality DNA can be prepared for downstream applications in <30 minutes.

语种: 英语
WOS记录号: WOS:000314886100008
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/50280
Appears in Collections:基础医学院_病理学系_期刊论文

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Deparaffinization and Lysis by Hydrothermal Pressure (Pressure Cooking) Coupled With Chaotropic Salt Column Purification A Rapid and Efficient Method of DNA Extraction From Formalin-fixed Paraffin-embedded Tissue.pdf(316KB)期刊论文出版稿限制开放 联系获取全文

作者单位: 1.Peking Univ, Hlth Sci Ctr, Dept Pathol, Beijing 100871, Peoples R China
2.Yale Univ, Sch Med, Dept Pathol, New Haven, CT 06520 USA

Recommended Citation:
Zhong, Haohao,Liu, Yan,Talmor, Monica,et al. Deparaffinization and Lysis by Hydrothermal Pressure (Pressure Cooking) Coupled With Chaotropic Salt Column Purification: A Rapid and Efficient Method of DNA Extraction From Formalin-fixed Paraffin-embedded Tissue[J]. DIAGNOSTIC MOLECULAR PATHOLOGY,2013,22(1):52-58.
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