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Int J Clin Exp Pathol. 2013;6(1):76-9. Epub 2012 Nov 20.

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

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  • 1Department of Radiobiology/Molecular Epidemiology, Radiation Effects Research Foundation 5-2 Hijiyama Park, Minami-ku, Hiroshima 732-0815, Japan. taga@rerf.or.jp

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 formalin-fixed, 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.

KEYWORDS:

DNA restoration; FFPE; PCR; lung tissue specimen

PMID:
23236545
[PubMed - indexed for MEDLINE]
PMCID:
PMC3515989
Free PMC Article
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