Enhanced detection of RNA from paraffin-embedded tissue using a panel of truncated gene-specific primers for reverse transcription

摘要

Formalin-fixed, paraffin-embedded tissues (PET) are a unique source of research material with the potential of providing biological information in conjunction with known clinical outcome. Specifically, these tissues could be an ideal resource for validating newly discovered genes as diagnostic and/or prognostic molecular markers in retrospective studies. Unfortunately, RNA isolated from PET samples is considered to be a poor material for molecular analyses, since RNA is frequently degraded to 100- to 200-bp fragments by endogenous and exogenous ribonucleases (RNase). In addition, Masuda et al. have shown that RNA. and in particular the poly(A) tail of messenger RNA (mRNA), is chemically modified, making it a poor template for cDNA synthesis.