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Isolation of functional RNA from cactus fruit

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Abstract

Isolating RNA from cactus fruit is notoriously difficult because the fruit contains high amounts of secondary metabolites and polysaccharides. These form insoluble complexes with nucleic acids during extraction and can inhibit enzyme action. Our procedure allows for the extraction of RNA from finely ground tissue. The RNA we isolated was of high quality and undegraded, as gauged by spectrophotometry and electrophoresis in agarose gels. Quality was further assessed through use of the RNA in RT-PCR and northern blot analysis, indicating that it could be used to construct cDNA libraries. Using this modified protocol, 90μg of RNA was routinely obtained from 1 g of dried cactus fruit. Isolating RNA from other polysaccharide-rich fruits was also possible.

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Abbreviations

DEPC:

diethyl pyrocarbonate

dNTP:

deoxyribonucleotide triphosphate

PG:

polygalacturonase

RT-PCR:

reverse transcription-polymerase chain reaction

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Authors and Affiliations

  1. Centro de Investigación y de Estudios Avanzados del IPN, Unidad Irapuato, Apartado Postal 629, 36500, Irapuato, Gto, México

    María Leonor Valderrama-Cháirez, Andrés Cruz-Hernández & Octavio Paredes-López

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  1. María Leonor Valderrama-Cháirez
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  2. Andrés Cruz-Hernández
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  3. Octavio Paredes-López
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Correspondence to Octavio Paredes-López.

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Valderrama-Cháirez, M.L., Cruz-Hernández, A. & Paredes-López, O. Isolation of functional RNA from cactus fruit. Plant Mol Biol Rep 20, 279–286 (2002). https://doi.org/10.1007/BF02782463

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  • DOI: https://doi.org/10.1007/BF02782463

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