RNA isolation

An alternative cetyltrimethylammonium bromide-based protocol for RNA isolation from blackberry (Rubus L.)

Q. Chen, Yu, H. W., Wang, X. R., Xie, X. L., Yue, X. Y., and Tang, H. R., An alternative cetyltrimethylammonium bromide-based protocol for RNA isolation from blackberry (Rubus L.), vol. 11. pp. 1773-1782, 2012.

Isolation of high-quality RNA free of contaminants, such as polyphenols, proteins, plant secondary metabolites, and genomic DNA from plant tissues, is usually a challenging but crucial step for molecular analysis. We developed a novel protocol based on the cetyltrimethylammonium bromide method to isolate high-quality RNA from blackberry plant tissues, especially fruits. Most DNA was removed when acetic acid was utilized, before RNA precipitation. Thus, lithium chloride, a reagent widely used for RNA purification, was not needed. The isolation time was shortened to less than 3 h.

Efficient isolation of high-quality RNA from lotus Nelumbo nucifera ssp nucifera tissues

Y. J. Zhang, Hao, X. Y., Liang, Z. S., Ke, W. D., and Guo, H. B., Efficient isolation of high-quality RNA from lotus Nelumbo nucifera ssp nucifera tissues, vol. 12. pp. 223-229, 2013.

Nelumbo nucifera is widely used as food, as an ornamental, in medicine, and as packing material; it is also reported to have anti-HIV effects and antioxidant capacity. We sought an improved method for extracting high-quality total RNA from different tissues of N. nucifera. Four methods for RNA extraction were assessed for their ability to recover high-quality RNA applicable for evaluation of polyphenol oxidase (PPO) gene expression profiles.

Isolation of retro-transcribed RNA from in vitro Mycosphaerella fijiensis-infected banana leaves

C. M. Rodríguez-García, Peraza-Echeverría, L., Islas-Flores, I. R., Canto-Canché, B. B., and Grijalva-Arango, R., Isolation of retro-transcribed RNA from in vitro Mycosphaerella fijiensis-infected banana leaves, vol. 9, pp. 1460-1468, 2010.

High polyphenol and polysaccharide levels in plant tissues such as banana fruit and leaves constitute a significant challenge to the extraction of sufficient amounts of high-quality RNA required for cDNA library synthesis and molecular analysis.

An optimized preparation method to obtain high-quality RNA from dry sunflower seeds

J. Yang and Ma, X. B., An optimized preparation method to obtain high-quality RNA from dry sunflower seeds, vol. 10, pp. 160-168, 2011.

In an attempt to isolate high-quality, intact total RNA from sunflower (Helianthus annuus) seeds for investigation of the molecular mechanisms of mutations, we tested various procedures, using kits, including RNAiso Plus, RNAiso Plus+RNAiso-mate for Plant Tissue, Trizol, and the Qi method, but no high-quality total RNA of high integrity was obtained with any of these methods, probably due to the high content of polyphenols, polysaccharides, and secondary metabolites in mature sunflower seeds. Modifications were made to the Qi method.

A rapid and efficient method for isolation of total RNA from Euglena gracilis (Euglenoidea)

D. González-Mendoza, Morales-Trejo, A., and Brito-Vera, H., A rapid and efficient method for isolation of total RNA from Euglena gracilis (Euglenoidea), vol. 8, pp. 482-486, 2009.

RNA isolation is essential to the study of gene expression at the molecular level. However, it is difficult to isolate RNA from organisms that contain large amounts of polysaccharides or other compounds that bind or coprecipitate with RNA, such as the unicellular protist Euglena gracilis. Currently, there is no commercial kit available that is specific for the isolation of high-quality RNA from this organism. Since it contains large amount of polysaccharides, the common protocols for RNA isolation usually result in poor yields when applied to E. gracilis.

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