Wednesday 7 December 2011
Molecular signatures of tumors with prognostic and predictive value are now available. However, several technical problems prevent the performing of gene expression profiles in routine diagnostics.
The highly sensitive fluorescence-based real-time quantitative reverse transcriptase-polymerase chain reaction procedure is able to analyze mRNA levels from formalin-fixed paraffin-embedded (FFPE) tissues, the most commonly available source of tumor samples with well-documented information.
However, the time-dependent fragmentation and small amount of RNA extracted from FFPE requires a preliminary mRNA amplification step to amplify small amounts of mRNA without significantly distorting relative mRNA levels.
Commercial cDNA preamplification kit
TaqMan PreAmp Master Mix Kit
Reliability and reproducibility of a RNA preamplification method for low-density array analysis from formalin-fixed paraffin-embedded breast cancer samples. Ciotti P, Garuti A, Ballestrero A, Cirmena G, Chiaramondia M, Baccini P, Bellone E, Mandich P. Diagn Mol Pathol. 2009 Jun;18(2):112-8. PMID: 19430292