Abstract
When conventional RNA isolation methods optimized for pine seedlings are applied to needles of adult pine trees, poor-quality RNA results. Here we describe a modified procedure to isolate high-quality RNA from needles of 30-year-old maritime pines, exhibiting high levels of phenolics, polysaccharides, and RNases. Major changes are the inclusion of proteinase K in the extraction medium followed by incubation at 42°C. Integrity and purity were evaluated by using denaturing gel electrophoresis and spectrophotometry (A260/A230 and A260/A280). The total RNA could be successfully used for poly(A)+-RNA isolation and cDNA library construction.
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Abbreviations
- CTAB:
-
hexadecyltrimethylammonium bromide
- DEPC:
-
diethyl pyrocarbonate
- FW:
-
fresh weight
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Azevedo, H., Lino-Neto, T. & Tavares, R.M. An improved method for high-quality RNA isolation from needles of adult maritime pine trees. Plant Mol Biol Rep 21, 333–338 (2003). https://doi.org/10.1007/BF02772582
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DOI: https://doi.org/10.1007/BF02772582