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

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
Kiyohiro HamataniKei Nakachi

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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Citations

Apr 22, 2008·BMC Medical Genomics·Andrea OberliRolf Jaggi
Feb 23, 2011·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Shan-Rong ShiClive R Taylor
Oct 12, 2012·Thyroid : Official Journal of the American Thyroid Association·Kiyohiro HamataniYoichiro Kusunoki
Sep 30, 2009·Thyroid : Official Journal of the American Thyroid Association·Kiyohiro HamataniKei Nakachi
Jan 21, 2016·Nature·Alexander J WestermannJörg Vogel
May 4, 2012·Growth Hormone & IGF Research : Official Journal of the Growth Hormone Research Society and the International IGF Research Society·Sean C HarringtonPaul Haluska
Feb 23, 2008·Experimental and Molecular Pathology·Melissa L CoxRobert W Dunstan
May 8, 2010·Veterinary Ophthalmology·Rodrigo Antonio LopesAlexandre Lima de Andrade
Sep 20, 2007·Journal of Nippon Medical School = Nippon Ika Daigaku Zasshi·Toshihiro TakizawaJohn M Robinson
Aug 12, 2014·BioMed Research International·Gisele Rodrigues GouveiaJuliana Pereira
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Apr 2, 2013·Oral Surgery, Oral Medicine, Oral Pathology and Oral Radiology·Jean-Luc C MougeotPeter B Lockhart
Oct 21, 2015·Biopreservation and Biobanking·Keisuke HaraEiji Uchida
May 27, 2010·Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B·Isabella DottiGiorgio Stanta

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Methods Mentioned

BETA
PCR
chemical modification
electrophoresis

Software Mentioned

1D Image Analysis

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