Abstract
A protocol is described for rapid RNA isolation from various plant species and tissues rich in polyphenolics and polysaccharides. The method is based on the Nucleon PhytoPure™ system without the use of phenol. The procedure can be completed within 1 h and many samples can be processed at the same time. The yield ranged from 240 μg up to 3 mg per gram of tissue with an average purity measured as A260/280 of 1.85. The RNA was of sufficient quality for use in RT-PCR reactions. Quantitation of single-stranded cDNA was carried out with the RiboGreen™ reagent and of PCR products with the PicoGreen™ reagent.
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Abbreviations
- CTAB:
-
cetyltrimethylammonium bromide
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Kiefer, E., Heller, W. & Ernst, D. A simple and efficient protocol for isolation of functional RNA from plant tissues rich in secondary metabolites. Plant Mol Biol Rep 18, 33–39 (2000). https://doi.org/10.1007/BF02825291
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DOI: https://doi.org/10.1007/BF02825291