Improved RT-PCR amplification for molecular analyses with long-term preserved formalin-fixed, paraffin-embedded tissue specimens

Hamatani K, Eguchi H, Takahashi K, Koyama K, Mukai M, Ito R, Taga M, Nakachi K
J Histochem Cytochem 54(7):773-80, 2006

Summary

Recently, in addition to DNA, RNA extracted from archival tissue specimens has become an invaluable source of material for molecular biological analysis. Successful amplification with PCR/RT-PCR is problematic when using amplicons of short size due to degradation of DNA or RNA. We established an improved method for efficient RT-PCR amplification of RNA extracted from archival formalin-fixed, paraffin-embedded tissue by the elimination of RNA modification and the restoration of RNA template activity. Namely, the preheating in citrate buffer (pH 4.0) of RNA extracted from long-term preserved tissue specimens resulted in significantly increased efficiency of RT-PCR.

Reproduced, with permission, from Hamatani K, Eguchi H, Takahashi K, Koyama K, Mukai M, Ito R, Taga M, Nakachi K: Improved RT-PCR amplification for molecular analyses with long-term preserved formalin-fixed, paraffin-embedded tissue specimens. Journal of Histochemistry & Cytochemistry 54(7):773-80, 2006