IJCEP Copyright © 2007-All rights reserved.
Int J Clin Exp Pathol 2013;6(1):76-79

Original Article
Improved PCR amplification for molecular analysis using DNA from long-term preserved
formalin-fixed, paraffin-embedded lung cancer tissue specimens

Masataka Taga*, Hidetaka Eguchi*, Tomoko Shinohara, Keiko Takahashi, Reiko Ito, Wataru Yasui, Kei
Nakachi, Yoichiro Kusunoki, Kiyohiro Hamatani

Department of Radiobiology/Molecular Epidemiology, Radiation Effects Research Foundation; Present address: Research Institute for
Development Therapeutics & Translational Research Center, International Medical Center, Saitama Medical University; Department of
Molecular Pathology, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan. *These authors contributed equally to
this work.

Received September 13, 2012; Accepted October 26, 2012; Epub November 20, 2012; Published January 1, 2013

Abstract: Archival tissue specimens are valuable resources of materials for molecular biological analyses in retrospective studies, especially
for rare diseases or those associated with exposure to uncommon environmental events. Although successful amplification with PCR is
essential for analysis of DNA extracted from archival formalinfixed, paraffin-embedded (FFPE) tissue specimens, we have often encountered
problems with poor PCR amplification of target fragments. To overcome this, we examined whether heat treatment in alkaline solution could
efficiently restore the PCR template activity of DNA that had already been extracted from FFPE lung cancer tissue specimens. The effect of the
heat treatment was assessed by PCR for the TP53 gene and other lung cancer-related gene loci. The heat treatment of DNA samples in borate
buffer resulted in successful PCR amplification of DNA fragments ranging from 91 to 152 bp. This technique for restoration of template activity
of DNA for PCR amplification is very simple and economical, and requires no special apparatus, so it may be applicable for molecular analysis
of DNA samples from FFPE tissue specimens at various laboratories. (IJCP1209008).

Keywords: FFPE, lung tissue specimen, DNA restoration, PCR

Address all correspondence to:
Dr. Masataka Taga
Department of Radiobiology/Molecular Epidemiology
Radiation Effects Research Foundation
5-2 Hijiyama Park, Minami-ku
Hiroshima 732-0815, Japan.
Tel: +81-82-261-3131; Fax: +81-82-261-3170
E-mail: taga@rerf.or.jp