Improved RT-PCR amplification for molecular analyses with long-term preserved formalin-fixed, paraffin-embedded tissue specimens. |
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Authors: | Kiyohiro Hamatani Hidetaka Eguchi Keiko Takahashi Kazuaki Koyama Mayumi Mukai Reiko Ito Masataka Taga Wataru Yasui Kei Nakachi |
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Affiliation: | Department of Radiobiology/Molecular Epidemiology, Radiation Effects Research Foundation, Hiroshima-shi, Hiroshima 732-0815, Japan. hamatani@rerf.or.jp |
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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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