A method for isolating total RNA from pear leaves |
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| Authors: | M Malnoy J P Reynoird F Mourgues E Chevreau P Simoneau |
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| Affiliation: | (1) INRA Unitée d’Améelioration des Espèces Fruitières et Ornementales, BP 57, 49071 Beaucouzée Cedex, France;(2) UMR 077 Pathologie Véegéetale, Facultée des sciences, 2bd Lavoisier, 49045 Angers Cedex, France;(3) Present address: Dept of Plant Pathology, Cornell University, 14456 Geneva, NY, USA;(4) Present address: Forli Section, Fruit Tree Research Station ISF, 2 via Punta di Ferro, CP 7178, 47100 Forli, Italy |
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| Abstract: | Isolation of high quality RNA fromRosaceae species is particularly difficult. These plants contain considerable amounts of plant polyphenolic compounds and polysaccharides
that copurify with RNA, often rendering it unsuitable for either cDNA synthesis and/or hybridization in northern analyses.
We describe a method for RNA isolation from pear leaves that is modified from that of Manning (1990). The procedure includes
i) an extraction with phenol and PVPP, to remove proteins and polyphenols ii) two purifications by LiCl, with a 2-butoxyethanol
treatment between the LiCl steps. The method results in high quality RNA suitable for RT-PCR and northern blot experiments. |
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| Keywords: | pear polysaccharide RNA contamination RNA extraction |
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