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dc.contributor.author
Jones, Leandro Roberto
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dc.contributor.author
Zandomeni, Rubén O.
dc.contributor.author
Weber, E. Laura
dc.date.available
2017-05-19T21:19:33Z
dc.date.issued
2006-12
dc.identifier.citation
Jones, 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.identifier.issn
0166-0934
dc.identifier.uri
http://hdl.handle.net/11336/16792
dc.description.abstract
A 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.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier Science Inc
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.subject.classification
Ciencias de las Plantas, Botánica
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
A long distance RT-PCR able to amplify the Pestivirus genome
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2017-05-17T13:52:06Z
dc.journal.volume
134
dc.journal.number
1-2
dc.journal.pagination
197-204
dc.journal.pais
Países Bajos
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dc.journal.ciudad
Amsterdam
dc.description.fil
Fil: 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.description.fil
Fil: Zandomeni, Rubén O.. Instituto Nacional de Tecnología Agropecuaria. Instituto de Microbiología y Zoología Agrícola; Argentina
dc.description.fil
Fil: Weber, E. Laura. Instituto Nacional de Tecnología Agropecuaria. Instituto de Virología; Argentina
dc.journal.title
Journal of Virological Methods
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.jviromet.2006.01.005
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0166093406000322
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