Isolation of high-quality total RNA from leaves of myrciaria dubia Camu camu
Myrciaria dubia is a main source of vitamin C for people in the Amazon region. Molecular studies of M. dubia require high-quality total RNA from different tissues. So far, no protocols have been reported for total RNA isolation from leaves of this species. The objective of this research was to devel...
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| Format: | info:eu-repo/semantics/article |
| Language: | Inglés |
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Taylor & Francis
2017
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| Online Access: | https://repositorio.inia.gob.pe/handle/20.500.12955/72 http://dx.doi.org/10.1080/10826068.2012.759968 |
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| author | Castro Gómez, Juan Carlos Egoavil Reátegui, Alina del Carmen Torres Flores, Julián Ramírez Saavedra, Roberson Imán Correa, Sixto Alfredo |
| author_browse | Castro Gómez, Juan Carlos Egoavil Reátegui, Alina del Carmen Imán Correa, Sixto Alfredo Ramírez Saavedra, Roberson Torres Flores, Julián |
| author_facet | Castro Gómez, Juan Carlos Egoavil Reátegui, Alina del Carmen Torres Flores, Julián Ramírez Saavedra, Roberson Imán Correa, Sixto Alfredo |
| author_sort | Castro Gómez, Juan Carlos |
| collection | Repositorio INIA |
| description | Myrciaria dubia is a main source of vitamin C for people in the Amazon region. Molecular studies of M. dubia require high-quality total RNA from different tissues. So far, no protocols have been reported for total RNA isolation from leaves of this species. The objective of this research was to develop protocols for extracting high-quality total RNA from leaves of M. dubia. Total RNA was purified following two modified protocols developed for leaves of other species (by Zeng and Yang, and by Reid et al.) and one modified protocol developed for fruits of the studied species (by Silva). Quantity and quality of purified total RNA were assessed by spectrophotometric and electrophoretic analysis. Additionally, quality of total RNA was evaluated with reverse-transcription polymerase chain reaction (RT-PCR). With these three modified protocols we were able to isolate high-quality RNA (A260nm/A280nm >1.9 and A260nm/A230nm >2.0). Highest yield was produced with the Zeng and Yang modified protocol (384+/-46microg ARN/g fresh weight). Furthermore, electrophoretic analysis showed the integrity of isolated RNA and the absence of DNA. Another proof of the high quality of our purified RNA was the successful cDNA synthesis and amplification of a segment of the M. dubia actin 1 gene. We report three modified protocols for isolation total RNA from leaves of M. dubia. The modified protocols are easy, rapid, low in cost, and effective for high-quality and quantity total RNA isolation suitable for cDNA synthesis and polymerase chain reaction. |
| format | info:eu-repo/semantics/article |
| id | INIA72 |
| institution | Institucional Nacional de Innovación Agraria |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| publisher | Taylor & Francis |
| publisherStr | Taylor & Francis |
| record_format | dspace |
| spelling | INIA722023-11-28T14:55:55Z Isolation of high-quality total RNA from leaves of myrciaria dubia Camu camu Castro Gómez, Juan Carlos Egoavil Reátegui, Alina del Carmen Torres Flores, Julián Ramírez Saavedra, Roberson Imán Correa, Sixto Alfredo Mejoramiento genético de plantas RNA ARN Frutas tropicales Ácido ascórbico Vitamina C Biotecnología Agrícola y de alimentos Myrciaria dubia is a main source of vitamin C for people in the Amazon region. Molecular studies of M. dubia require high-quality total RNA from different tissues. So far, no protocols have been reported for total RNA isolation from leaves of this species. The objective of this research was to develop protocols for extracting high-quality total RNA from leaves of M. dubia. Total RNA was purified following two modified protocols developed for leaves of other species (by Zeng and Yang, and by Reid et al.) and one modified protocol developed for fruits of the studied species (by Silva). Quantity and quality of purified total RNA were assessed by spectrophotometric and electrophoretic analysis. Additionally, quality of total RNA was evaluated with reverse-transcription polymerase chain reaction (RT-PCR). With these three modified protocols we were able to isolate high-quality RNA (A260nm/A280nm >1.9 and A260nm/A230nm >2.0). Highest yield was produced with the Zeng and Yang modified protocol (384+/-46microg ARN/g fresh weight). Furthermore, electrophoretic analysis showed the integrity of isolated RNA and the absence of DNA. Another proof of the high quality of our purified RNA was the successful cDNA synthesis and amplification of a segment of the M. dubia actin 1 gene. We report three modified protocols for isolation total RNA from leaves of M. dubia. The modified protocols are easy, rapid, low in cost, and effective for high-quality and quantity total RNA isolation suitable for cDNA synthesis and polymerase chain reaction. Peer reviewed 2017-11-12T18:30:18Z 2017-11-12T18:30:18Z 2013-08-18 info:eu-repo/semantics/article Castro, J. C.; Egoavil, A.D.C.; Torres, J.; Ramírez, R.; Cobos, M. & Imán, S. A. (2013). Isolation of high-quality total RNA from leaves of myrciaria dubia Camu Camu. Preparative Biochemistry and Biotechnology, 43(6), 527–538. doi: 10.1080/10826068.2012.759968 10826068 https://repositorio.inia.gob.pe/handle/20.500.12955/72 Preparative Biochemistry and Biotechnology http://dx.doi.org/10.1080/10826068.2012.759968 eng Preparative Biochemistry and Biotechnology, v.43, n.6, 2013, pp.527–538 http://dx.doi.org/10.1080/10826068.2012.759968 info:eu-repo/semantics/restrictedAccess application/pdf application/pdf Taylor & Francis Reino Unido Instituto Nacional de Innovación Agraria Repositorio Institucional - INIA |
| spellingShingle | Mejoramiento genético de plantas RNA ARN Frutas tropicales Ácido ascórbico Vitamina C Biotecnología Agrícola y de alimentos Castro Gómez, Juan Carlos Egoavil Reátegui, Alina del Carmen Torres Flores, Julián Ramírez Saavedra, Roberson Imán Correa, Sixto Alfredo Isolation of high-quality total RNA from leaves of myrciaria dubia Camu camu |
| title | Isolation of high-quality total RNA from leaves of myrciaria dubia Camu camu |
| title_full | Isolation of high-quality total RNA from leaves of myrciaria dubia Camu camu |
| title_fullStr | Isolation of high-quality total RNA from leaves of myrciaria dubia Camu camu |
| title_full_unstemmed | Isolation of high-quality total RNA from leaves of myrciaria dubia Camu camu |
| title_short | Isolation of high-quality total RNA from leaves of myrciaria dubia Camu camu |
| title_sort | isolation of high quality total rna from leaves of myrciaria dubia camu camu |
| topic | Mejoramiento genético de plantas RNA ARN Frutas tropicales Ácido ascórbico Vitamina C Biotecnología Agrícola y de alimentos |
| url | https://repositorio.inia.gob.pe/handle/20.500.12955/72 http://dx.doi.org/10.1080/10826068.2012.759968 |
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