Comparação de três protocolos distintos para extração de RNA de amostras fixadas em formalina e emblocadas em parafina

Abstract

Introduction: Formalin fixed paraffin embedded (FFPE) tissues are an important sample source for retrospective studies. Despite its ability to preserve proteins and cell morphology, formalin hinders Molecular Biology tests once it fragments and chemically modifies nucleic acids, particularly RNA. Objective: To compare the efficiency of three different RNA extraction protocols for gene expression analysis of FFEP tissues. Material and methods: RNA was extracted from FFPE samples of human lymph by means of Ambion and Arcturus Bioscience kits and the classical Trizol method. After extraction, RNA was quantified and tested for amplification through real time polymerase chain reaction (RT-PCR) using glyceraldehydes-3 phosphate dehydrogenase (GAPDH) endogenous gene primers. Discussion/conclusion: All the protocols produced sufficient and adequate amounts of total RNA. However, only protocols using Arcturus and Ambion kits generated suitable RNA for PCR amplification.