A pressure cooking-based DNA extraction from archival formalin-fixed, paraffin-embedded tissue

作者：Chung Joon Yong; Yi Joo Mi; Xie Ran; Brown Victoria; Lee Olivia; Ahuja Nita; Braunschweig Till; Hewitt Stephen M^*

来源：Analytical Biochemistry, 2012, 425(2): 128-134.

DOI：10.1016/j.ab.2012.03.012

• DNA extraction
• DNA methylation
• Methylation-specific PCR (MSP)
• Formalin-fixed paraffin-embedded tissues
• Intraepithelial neoplasia

As emerging novel DNA-based methodologies are adopted, nucleic acid-based assays depend critically on the quality and quantity of extracted DNA. Formalin-fixed, paraffin embedded (FFPE) tissue samples provide an invaluable resource for subsequent molecular studies of clinical phenotypes, but high-quality DNA extraction from archival FFPE tissue specimens remains complex and time-consuming. To address this challenge, we have developed a reliable rapid DNA extraction method for FFPE tissue specimens. It is based on deparaffinization at high temperature coupled with relieving crosslink in a pressure cooker. The DNA yield by this rapid method resulted in an average 1.8-fold increase in comparison with the commercial kit and OD 260/280 ratios between 1.87 and 1.95. The DNA obtained by the rapid method was suitable for methylation analyses in colon cancer patients. These data suggest that this new DNA extraction method coupled with methylation-specific polymerase chain reaction can be used for epigenetic studies with the advantages of rapidity and high quality and may contribute to the development of biomarkers in clinical studies. Published by Elsevier Inc.

• 出版日期2012-6-15