Optimized Method for High‐Quality RNA Extraction from Formalin‐Fixed Paraffin‐Embedded Tissues

Abstract

AbstractRNA extraction from formalin‐fixed paraffin‐embedded tissues (FFPE) presents a notable challenge. Extracting RNA from FFPE tissues has been challenging due to RNA degradation and chemical bonding between RNA and proteins during the fixation process. This results in reduced concentration and quality of the extracted RNA from FFPE tissues, impacting subsequent downstream analyses. In this study, we present an optimized approach involves replacing the tissue temperature‐digestion step with tissue crushing and homogenization. RNA extraction was performed on 100 FFPE tissue blocks obtained from the archive of Pathology Department of Shiraz University of Medical Sciences. After deparafinization of the tissues, RNA was extracted using this modified method, and the results compared to those of a standard extraction procedure and commercial kits used for FFPE tissue. Then gel electrophoresis and Real time PCR were done to check the quality and integrity of the extracted RNA. The results show that the concentration of RNA obtained from this method was significantly (P value <0.0001) higher than the other methods. In conclusion, we have successfully introduced a modified method that enables the extraction of high‐quality and intact RNA from FFPE tissue. This method stands as a reliable option for molecular biology studies necessitating precise RNA extraction.