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. 2006 Jul;54(7):773-80.
doi: 10.1369/jhc.5A6859.2006. Epub 2006 Mar 3.

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

Kiyohiro Hamatani  1 , Hidetaka Eguchi, Keiko Takahashi, Kazuaki Koyama, Mayumi Mukai, Reiko Ito, Masataka Taga, Wataru Yasui, Kei Nakachi
Affiliations

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

Kiyohiro Hamatani et al. J Histochem Cytochem. 2006 Jul.

Abstract

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.

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