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

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• 作者：Herlander Azevedo ; Teresa Lino-Neto and Rui M. Tavares
• 关键词：cDNA library construction ; maritime pine needles ; RNA isolation
• 刊名：Plant Molecular Biology Reporter
• 出版年：2003
• 出版时间：December, 2003
• 年：2003
• 卷：21
• 期：4
• 页码：333-338
• 全文大小：354.5 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Plant Sciences
  Plant Physiology
  
• 出版者：Springer Netherlands
• ISSN：1572-9818

文摘

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.