University of Hertfordshire Research Archive

        JavaScript is disabled for your browser. Some features of this site may not work without it.

        Browse

        All of UHRABy Issue DateAuthorsTitlesThis CollectionBy Issue DateAuthorsTitles

        Arkivum Files

        My Downloads
        View Item 
        • UHRA Home
        • University of Hertfordshire
        • Research publications
        • View Item
        • UHRA Home
        • University of Hertfordshire
        • Research publications
        • View Item

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

        View/Open
        902845.pdf (PDF, 260Kb)
        Author
        Witchell, J.
        Varshney, D.
        Gajjar, T.
        Wangoo, A.
        Goyal, M.
        Attention
        2299/2843
        Abstract
        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.
        Publication date
        2008
        Published in
        Pathology, Research and Practice
        Published version
        https://doi.org/10.1016/j.prp.2007.09.002
        Other links
        http://hdl.handle.net/2299/2843
        Metadata
        Show full item record
        Keep in touch

        © 2019 University of Hertfordshire

        I want to...

        • Apply for a course
        • Download a Prospectus
        • Find a job at the University
        • Make a complaint
        • Contact the Press Office

        Go to...

        • Accommodation booking
        • Your student record
        • Bayfordbury
        • KASPAR
        • UH Arts

        The small print

        • Terms of use
        • Privacy and cookies
        • Criminal Finances Act 2017
        • Modern Slavery Act 2015
        • Sitemap

        Find/Contact us

        • T: +44 (0)1707 284000
        • E: ask@herts.ac.uk
        • Where to find us
        • Parking
        • hr
        • qaa
        • stonewall
        • AMBA
        • ECU Race Charter
        • disability confident
        • AthenaSwan