Examinando por Materia "ARN"

Mostrando 1 - 3 de 3

Isolation of high-quality total RNA from leaves of myrciaria dubia Camu camu
(Taylor & Francis, 2013-08-18) Castro Gómez, Juan Carlos; Egoavil Reátegui, Alina del Carmen; Torres Flores, Julián; Ramírez Saavedra, Roberson; Imán Correa, Sixto Alfredo
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.
Manual de caracterización genotípica de recursos genéticos
(Instituto Nacional de Innovación Agraria, 2021-03) Biondi Thorndike, Jorge Andrés; Fernández Huaytalla, Elizabeth; Zorrilla Cisneros, Cinthya; Amasifuen Guerra, Carlos Alberto
El Ministerio de Desarrollo Agrario y Riego (MIDAGRI) a través del Instituto Nacional de Innovación Agraria (INIA), pone a disposición el presente manual titulado “Caracterización genotípica de recursos genéticos”, donde se describen los diferentes métodos y procedimientos utilizados para el proceso de colecta de muestras, extracción de ácidos nucleicos, obtención de marcadores moleculares y diagnóstico del estado fitosanitario; realizados exitosamente en los cultivos de yuca, oca, tomate, arracacha y yacón. El presente manual es el resultado de numerosos estudios sobre la diversidad genética y caracterización molecular de las colecciones de germoplasma, ubicadas estratégicamente en el territorio nacional a través de las Estaciones Experimentales Agrarias (EEAs) del INIA.
Myrciaria dubia “Camu Camu” Fruit: Health-Promoting Phytochemicals and Functional Genomic Characteristics
(IntechOpen, 2018-06-13) Castro Gómez, Juan Carlos; Maddox, J. Dylan; Cobos Ruiz, Marianela; Imán Correa, Sixto Alfredo
Camu camu is a typical Amazon native fruit shrub that possesses a diploid genome, moderate genetic diversity, and population structure. The fruits accumulate several essential nutrients and synthesize L-ascorbic acid (vitamin C) in great quantities and an array of diverse secondary metabolites with corroborated in vitro and in vivo health-promoting activities. These beneficial effects include antioxidative and antiinflammatory activities, antiobesity, hypolipidemic, antihypertensive and antidiabetic effects, DNA damage and cancer protection effects, and other bioactivities. Many health-promoting phytochemicals are biosynthesized in several metabolic pathways of camu camu. Their reconstruction from the fruit transcriptome database was accomplished by our research group. These include basic metabolic pathways such as glycolysis and pentose phosphate pathway, vitamin C biosynthesis pathways, and pathways involved in secondary metabolites production. Due to their agronomic potential and fruits growing demand, recently, based on an ideotype, programs were initiated for their domestication and genetic improvement, but so far with very negligible achievements. Consequently, we propose new strategies to accelerate the processes of domestication and genetic improvement based on state of the art technologies for multiomic data analysis and innovative molecular tools.