Wednesday 7 December 2011
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