An Improved Protocol for Extraction and RT-PCR Detection of Citrus Viroids
The great sensitivity of RT-PCR makes this technique a desirable alternative to other diagnostic methods for detection of viroids. In previous studies, difficulties have been encountered in obtaining amplicons from samples known to contain viroids. The aim of the present study was to minimize extrac...
| Autores principales: | , |
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| Formato: | conferenceObject |
| Lenguaje: | Inglés |
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IOCV
2021
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.11939/7703 https://iocv.ucr.edu/sites/g/files/rcwecm4696/files/2020-06/452-455%20IOCV%2016.pdf |
| _version_ | 1855032654686060544 |
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| author | Bernad, Lucia Durán-Vila, Núria |
| author2 | Hilf, M. E. |
| author_browse | Bernad, Lucia Durán-Vila, Núria Hilf, M. E. |
| author_facet | Hilf, M. E. Bernad, Lucia Durán-Vila, Núria |
| author_sort | Bernad, Lucia |
| collection | ReDivia |
| description | The great sensitivity of RT-PCR makes this technique a desirable alternative to other diagnostic methods for detection of viroids. In previous studies, difficulties have been encountered in obtaining amplicons from samples known to contain viroids. The aim of the present study was to minimize extraction procedures and to design RT-PCR protocols based on the use of thermostable reverse transcriptases. The use of 27-nucleotide primers and reverse transcription at 60°C minimized renaturation of the viroid template and consistently yielded the expected PCR products for each viroid. The protocol has been tested with samples from a large number of viroid-host combinations grown under greenhouse conditions as well as with samples from the field. |
| format | conferenceObject |
| id | ReDivia7703 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | IOCV |
| publisherStr | IOCV |
| record_format | dspace |
| spelling | ReDivia77032025-04-25T14:53:09Z An Improved Protocol for Extraction and RT-PCR Detection of Citrus Viroids Bernad, Lucia Durán-Vila, Núria Hilf, M. E. RT-PCR Diagnosis methods Viroids Thermostable reverse transcriptases Citrus viroids H20 Plant diseases U30 Research methods The great sensitivity of RT-PCR makes this technique a desirable alternative to other diagnostic methods for detection of viroids. In previous studies, difficulties have been encountered in obtaining amplicons from samples known to contain viroids. The aim of the present study was to minimize extraction procedures and to design RT-PCR protocols based on the use of thermostable reverse transcriptases. The use of 27-nucleotide primers and reverse transcription at 60°C minimized renaturation of the viroid template and consistently yielded the expected PCR products for each viroid. The protocol has been tested with samples from a large number of viroid-host combinations grown under greenhouse conditions as well as with samples from the field. 2021-10-27T14:41:59Z 2021-10-27T14:41:59Z 2005 conferenceObject Bernad, L. & Durán-Vila, N. (2005). An Improved Protocol for Extraction and RT-PCR Detection of Citrus Viroids. Proceedings of the Sixteenth Conference of the International Organization of Citrus Virologists, 452-455. 2313-5123 0074-7203 (e-ISSN) http://hdl.handle.net/20.500.11939/7703 https://iocv.ucr.edu/sites/g/files/rcwecm4696/files/2020-06/452-455%20IOCV%2016.pdf en 2004-11-07 Sixteenth Conference of the International Organization of Citrus Virologists Monterrey, Mexico Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess IOCV electronico |
| spellingShingle | RT-PCR Diagnosis methods Viroids Thermostable reverse transcriptases Citrus viroids H20 Plant diseases U30 Research methods Bernad, Lucia Durán-Vila, Núria An Improved Protocol for Extraction and RT-PCR Detection of Citrus Viroids |
| title | An Improved Protocol for Extraction and RT-PCR Detection of Citrus Viroids |
| title_full | An Improved Protocol for Extraction and RT-PCR Detection of Citrus Viroids |
| title_fullStr | An Improved Protocol for Extraction and RT-PCR Detection of Citrus Viroids |
| title_full_unstemmed | An Improved Protocol for Extraction and RT-PCR Detection of Citrus Viroids |
| title_short | An Improved Protocol for Extraction and RT-PCR Detection of Citrus Viroids |
| title_sort | improved protocol for extraction and rt pcr detection of citrus viroids |
| topic | RT-PCR Diagnosis methods Viroids Thermostable reverse transcriptases Citrus viroids H20 Plant diseases U30 Research methods |
| url | http://hdl.handle.net/20.500.11939/7703 https://iocv.ucr.edu/sites/g/files/rcwecm4696/files/2020-06/452-455%20IOCV%2016.pdf |
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