A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA
Reverse transcription and polymerase chain reaction (RT-PCR) are being used increasingly for detection and typing RNA viruses. For this purpose, metal block thermal cyclers (MBTC) are considered to provide higher DNA yield, whereas air thermal cyclers (ATC) allow PCR amplification in a much shorter...
| Main Authors: | , , , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
2017
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| Online Access: | http://hdl.handle.net/20.500.11939/4478 |
| _version_ | 1855491761250500608 |
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| author | Sambade, A. Martin, Susana Olmos, Antonio Garcia, M. L. Cambra, Mariano Grau, Oscar Guerri, José Moreno, Pedro |
| author_browse | Cambra, Mariano Garcia, M. L. Grau, Oscar Guerri, José Martin, Susana Moreno, Pedro Olmos, Antonio Sambade, A. |
| author_facet | Sambade, A. Martin, Susana Olmos, Antonio Garcia, M. L. Cambra, Mariano Grau, Oscar Guerri, José Moreno, Pedro |
| author_sort | Sambade, A. |
| collection | ReDivia |
| description | Reverse transcription and polymerase chain reaction (RT-PCR) are being used increasingly for detection and typing RNA viruses. For this purpose, metal block thermal cyclers (MBTC) are considered to provide higher DNA yield, whereas air thermal cyclers (ATC) allow PCR amplification in a much shorter time. A fast ATC protocol (0 a denaturation, 0 s annealing, and 4-8 s elongation) was developed to amplify genomic segments from two RNA viruses, which allowed increasing the number of cycles without a parallel increase of non-specific DNA fragments. Under these conditions, 80-90 cycles with the ATC provided a DNA yield close to that of a standard 40-cycles MBTC protocol in about half the time. The DNA synthesised by the new procedure was highly specific and could be cloned readily. |
| format | Artículo |
| id | ReDivia4478 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia44782025-04-25T14:43:38Z A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA Sambade, A. Martin, Susana Olmos, Antonio Garcia, M. L. Cambra, Mariano Grau, Oscar Guerri, José Moreno, Pedro Reverse transcription and polymerase chain reaction (RT-PCR) are being used increasingly for detection and typing RNA viruses. For this purpose, metal block thermal cyclers (MBTC) are considered to provide higher DNA yield, whereas air thermal cyclers (ATC) allow PCR amplification in a much shorter time. A fast ATC protocol (0 a denaturation, 0 s annealing, and 4-8 s elongation) was developed to amplify genomic segments from two RNA viruses, which allowed increasing the number of cycles without a parallel increase of non-specific DNA fragments. Under these conditions, 80-90 cycles with the ATC provided a DNA yield close to that of a standard 40-cycles MBTC protocol in about half the time. The DNA synthesised by the new procedure was highly specific and could be cloned readily. 2017-06-01T10:10:14Z 2017-06-01T10:10:14Z 2000 JUN 2000 article Sambade, A., Martin, S., Olmos, A., Garcia, M.L., Cambra, M., Grau, O., Guerri, J., Moreno, P. (2000). A fast one-step reverse transcription and polymerase chain reaction (RT-PCR), amplification procedure providing highly specific complementary DNA from plant virus RNA. Journal of virological methods, 87(1-2), 25-28. 0166-0934 http://hdl.handle.net/20.500.11939/4478 10.1016/S0166-0934(00)00145-2 en openAccess Impreso |
| spellingShingle | Sambade, A. Martin, Susana Olmos, Antonio Garcia, M. L. Cambra, Mariano Grau, Oscar Guerri, José Moreno, Pedro A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA |
| title | A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA |
| title_full | A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA |
| title_fullStr | A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA |
| title_full_unstemmed | A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA |
| title_short | A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA |
| title_sort | fast one step reverse transcription and polymerase chain reaction rt pcr amplification procedure providing highly specific complementary dna from plant virus rna |
| url | http://hdl.handle.net/20.500.11939/4478 |
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