dc.contributor.author | Witchell, J. | |
dc.contributor.author | Varshney, D. | |
dc.contributor.author | Gajjar, T. | |
dc.contributor.author | Wangoo, A. | |
dc.contributor.author | Goyal, M. | |
dc.date.accessioned | 2009-01-26T13:21:04Z | |
dc.date.available | 2009-01-26T13:21:04Z | |
dc.date.issued | 2008 | |
dc.identifier.citation | Witchell , J , Varshney , D , Gajjar , T , Wangoo , A & Goyal , M 2008 , ' RNA isolation and quantitative PCR from HOPE- and formalin-fixed bovine lymph node tissues ' , Pathology, Research and Practice , vol. 204 , no. 2 , pp. 105-111 . https://doi.org/10.1016/j.prp.2007.09.002 | |
dc.identifier.issn | 0344-0338 | |
dc.identifier.other | dspace: 2299/2843 | |
dc.identifier.uri | http://hdl.handle.net/2299/2843 | |
dc.description | Original article can be found at: http://www.sciencedirect.com/science/journal/03440338 Copyright Elsevier GmbH. DOI: 10.1016/j.prp.2007.09.002 | |
dc.description.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. | en |
dc.format.extent | 266524 | |
dc.language.iso | eng | |
dc.relation.ispartof | Pathology, Research and Practice | |
dc.subject | Quantitative polymerase chain reaction | |
dc.subject | RNA extraction | |
dc.title | RNA isolation and quantitative PCR from HOPE- and formalin-fixed bovine lymph node tissues | en |
dc.contributor.institution | School of Life and Medical Sciences | |
dc.contributor.institution | Veterinary Science | |
dc.contributor.institution | Microbiology and Biotechnology | |
dc.contributor.institution | Biosciences Research Group | |
dc.contributor.institution | Centre for Research in Mechanisms of Disease and Drug Discovery | |
dc.contributor.institution | Department of Clinical, Pharmaceutical and Biological Science | |
dc.contributor.institution | Centre for Future Societies Research | |
dc.description.status | Peer reviewed | |
dc.identifier.url | http://www.scopus.com/inward/record.url?scp=38849088854&partnerID=8YFLogxK | |
rioxxterms.versionofrecord | 10.1016/j.prp.2007.09.002 | |
rioxxterms.type | Journal Article/Review | |
herts.preservation.rarelyaccessed | true | |