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dc.creatorJones, Leandro Roberto-
dc.creatorZandomeni, Rubén O.-
dc.creatorWeber, E. Laura-
dc.date2017-05-19T21:19:33Z-
dc.date2017-05-19T21:19:33Z-
dc.date2006-12-
dc.date2017-05-17T13:52:06Z-
dc.date.accessioned2019-04-29T15:43:56Z-
dc.date.available2019-04-29T15:43:56Z-
dc.date.issued2017-05-19T21:19:33Z-
dc.date.issued2017-05-19T21:19:33Z-
dc.date.issued2006-12-
dc.date.issued2017-05-17T13:52:06Z-
dc.identifierJones, Leandro Roberto; Zandomeni, Rubén O.; Weber, E. Laura; A long distance RT-PCR able to amplify the Pestivirus genome; Elsevier Science Inc; Journal of Virological Methods; 134; 1-2; 12-2006; 197-204-
dc.identifier0166-0934-
dc.identifierhttp://hdl.handle.net/11336/16792-
dc.identifier.urihttp://rodna.bn.gov.ar:8080/jspui/handle/bnmm/300569-
dc.descriptionA method to amplify long genomic regions (up to ∼ 12.3 kb) from pestiviruses in one RT-PCR is described. The difficulty in designing conserved Pestivirus primers for the amplification of genomes from highly divergent isolates simply by means of overlapping segments is demonstrated using new bioinformatic tools. An alternative procedure consisting of optimizing the length of the genomic cDNA fragments and their subsequent amplification by polymerase chain reaction (PCR) using a limited set of specific primers is described. The amplification of long DNA fragments from a variety of sources, including genomic, mitochondrial, and viral DNAs as well as cDNA produced by reverse transcription (RT) has been achieved using this methodology, known as long distance PCR. In the case of viruses, it is necessary to obtain viral particles from infected cells prior to RT procedures. This work provides improvements in four steps of long distance RT-PCR (L-RT-PCR): (i) preparation of a viral stock, (ii) preparation of template RNA, (iii) reverse transcription and (iv) amplification of the cDNA by LD-PCR. The usefulness of L-RT-PCR is discussed in the light of current knowledge on pestivirus diversity. The genomic sequence of Singer Arg reference strain obtained using this method is presented and characterized.-
dc.descriptionFil: Jones, Leandro Roberto. Universidad Nacional de la Patagonia Austral. Centro de Investigaciones y Transferencia Golfo San Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia Golfo San Jorge. Universidad Nacional de la Patagonia "san Juan Bosco". Centro de Investigaciones y Transferencia Golfo San Jorge; Argentina-
dc.descriptionFil: Zandomeni, Rubén O.. Instituto Nacional de Tecnología Agropecuaria. Instituto de Microbiología y Zoología Agrícola; Argentina-
dc.descriptionFil: Weber, E. Laura. Instituto Nacional de Tecnología Agropecuaria. Instituto de Virología; Argentina-
dc.formatapplication/pdf-
dc.formatapplication/pdf-
dc.languageeng-
dc.publisherElsevier Science Inc-
dc.relationinfo:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.jviromet.2006.01.005-
dc.relationinfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0166093406000322-
dc.rightsinfo:eu-repo/semantics/restrictedAccess-
dc.rightshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/-
dc.sourcereponame:CONICET Digital (CONICET)-
dc.sourceinstname:Consejo Nacional de Investigaciones Científicas y Técnicas-
dc.sourceinstacron:CONICET-
dc.subjectCiencias de las Plantas, Botánica-
dc.subjectCiencias Biológicas-
dc.subjectCIENCIAS NATURALES Y EXACTAS-
dc.titleA long distance RT-PCR able to amplify the Pestivirus genome-
dc.typeinfo:eu-repo/semantics/article-
dc.typeinfo:eu-repo/semantics/publishedVersion-
dc.typeinfo:ar-repo/semantics/articulo-
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