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学科主题基础医学
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
Zhong, Haohao1; Liu, Yan1; Talmor, Monica2; Wu, Bingquan1; Hui, Pei2
关键词hydrothermal pressure DNA extraction FFPE
刊名DIAGNOSTIC MOLECULAR PATHOLOGY
2013-03-01
DOI10.1097/PDM.0b013e318263f092
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
引用统计
被引频次:6[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/50280
专题北京大学基础医学院_病理学系
北京大学基础医学院
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
推荐引用方式
GB/T 7714
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.
APA Zhong, Haohao,Liu, Yan,Talmor, Monica,Wu, Bingquan,&Hui, Pei.(2013).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.DIAGNOSTIC MOLECULAR PATHOLOGY,22(1),52-58.
MLA Zhong, Haohao,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".DIAGNOSTIC MOLECULAR PATHOLOGY 22.1(2013):52-58.
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