An improved method for high-quality RNA isolation from needles of adult maritime pine trees

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, polysaccharide...

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Bibliographic Details
Published in:Plant molecular biology reporter 2003-12, Vol.21 (4), p.333-338
Main Authors: Azevedo, Herlânder, Lino-Neto, Teresa, Tavares, Rui M.
Format: Article
Language:English
Subjects:
Endopeptidase K
Gel electrophoresis
Phenols
Pine
Pine needles
Pine trees
Polyadenine
Polysaccharides
Proteinase
Ribonucleic acid
RNA
Saccharides
Seedlings
Spectrophotometry
Trees
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Summary: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 (A^sub 260^/A^sub 230^ and A^sub 260^/A^sub 280^). The total RNA could be successfully used for poly(A)^sup +^-RNA isolation and cDNA library construction.
ISSN:0735-9640
1572-9818
DOI:10.1007/BF02772582