RNA isolation and quantitative PCR from HOPE- and formalin-fixed bovine lymph node tissues

J. Witchell, D. Varshney, T. Gajjar, A. Wangoo, M. Goyal

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14 Citations (Scopus)
201 Downloads (Pure)


The use of RNA extracted from HOPE fixed tissues in quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is fairly novel. We compared qRT-PCR analysis of formalin and HOPE fixed, paraffin embedded lymph node tissues from M.bovis infected cattle, by extracting total RNA using a commercial kit (Ambion) and a trizol method. RNA extracted from HOPE fixed tissues showed comparable quantities between the commercial kit (82.7-107.9 μg/ml total RNA) and the trizol method (87-161.1 μg/ml total RNA), displaying a high degree of integrity when analysed by electrophoresis. RNA extracted from formalin fixed tissues using the commercial kit produced similar concentrations (80.6-145.7 μg/ml total RNA) in comparison to the HOPE tissue however the integrity was compromised. Extraction of RNA from the formalin fixed tissues using trizol was unsuccessful. Following qRT-PCR for GAPDH, total RNA from HOPE fixed tissues showed higher levels of target mRNA (4.05x10-2 pg/100ng total RNA using the commercial kit and 6.45x10-2 pg/100ng total RNA using trizol) in comparison to formalin fixed tissues (5.69x10-4 pg/100ng total RNA). This could be attributed to RNA degradation by exposure to formalin fixation. In conclusion, the HOPE fixative proved to be a better source for RNA extraction from cattle lymph nodes and subsequent qRT-PCR.
Original languageEnglish
Pages (from-to)105-111
JournalPathology, Research and Practice
Issue number2
Publication statusPublished - 2008


  • Quantitative polymerase chain reaction
  • RNA extraction


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