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dc.contributor.authorBernad, Lucia
dc.contributor.authorDurán-Vila, Núria
dc.date.accessioned2017-06-01T10:11:06Z
dc.date.available2017-06-01T10:11:06Z
dc.date.issued2006
dc.identifier.citationBernad, L., Duran-Vila, N. (2006). A novel RT-PCR approach for detection and characterization of citrus viroids. Molecular and cellular probes, 20(2), 105-113.
dc.identifier.issn0890-8508
dc.identifier.urihttp://hdl.handle.net/20.500.11939/4825
dc.description.abstractCitrus plants are natural hosts of five viroid species and a large number of sequence variants. Because of their small size, viroids lend themselves to various RT-PCR approaches for their detection and further characterization. The one-step RT-PCR approach proposed here is based on the synthesis of viroid-cDNA by reverse transcription at 60 degrees C using a viroid specific 27-mer primer followed by standard second strand synthesis plus PCR amplification with various primer pairs. According to the primers used, full or partial length viroid-DNA is obtained. The technique avoids amplicon contamination in routine diagnosis. The Suitability of the technique has been demonstrated using several nucleic acid extraction procedures and different viroid infected host species. The homogenization of tissue inside scaled plastic bags followed by nucleic acid extraction using a SDS/potassium acetate method is recommended because of its efficiency, simplicity and low cost. This extraction procedure, when coupled to the one-step RT-PCR approach, can be useful to avoid cross-contamination during routine diagnosis. A PCR strategy capable of discriminating between mild and severe strains of CEVd and identifying cachexia-inducing isolates of HSVd. is also described.
dc.language.isoen
dc.titleA novel RT-PCR approach for detection and characterization of citrus viroidsen
dc.typearticle
dc.authorAddressInstituto Valenciano de Investigaciones Agrarias (IVIA), Carretera CV-315, Km. 10’7, 46113 Moncada (Valencia), Españaes
dc.date.issuedFreeFormAPR 2006
dc.entidadIVIACentro de Protección Vegetal y Biotecnología
dc.identifier.doi10.1016/j.mcp.2005.11.001
dc.identifier.urlhttps://www.sciencedirect.com/science/article/abs/pii/S0890850805000964?via%3Dihub
dc.journal.abbreviatedTitleMol.Cell.Probes
dc.journal.issueNumber2
dc.journal.titleMolecular and cellular probes
dc.journal.volumeNumber20
dc.page.final113
dc.page.initial105
dc.rights.accessRightsopenAccess
dc.source.typeImpreso


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