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JHC exPRESS: First Published March 3, 2006. doi:10.1369/jhc.5A6859.2006
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A more recent version of this article appeared on July 1, 2006.
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Improved RT-PCR Amplification for Molecular Analyses with Long-term Preserved Formalin-fixed, Paraffin-embedded Tissue Specimens

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

1 Department of Radiobiology/Molecular Epidemiology, Radiation Effects Research Foundation, Hiroshima, Japan (KH,HE,KT,KK,MM,RI,MT,KN), and Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan (WY)

* To whom correspondence should be addressed. E-mail: hamatani{at}rerf.or.jp.

Submitted on October 19, 2005
Accepted on 14 February 2006


  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.

Key Words: preheating of RNA, archival formalin-fixed and paraffin-embedded tissue, citrate buffer, modification of RNA by formalin, RT-PCR amplification


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