Running head: RNA isolation from adult maritime pine needles Article type: Commentary Title: An improved method for high quality RNA isolation from needles of adult maritime pine trees

نویسندگان

  • HERLÂNDER AZEVEDO
  • TERESA LINO-NETO
  • RUI M. TAVARES
  • Rui M. Tavares
چکیده

Conventional RNA isolation methods optimised for pine seedlings have been shown to produce poor quality RNA when applied to needles of adult pine trees. We describe here a modified procedure to isolate high pure RNA from needles of thirty-year-old maritime pines, exhibiting high levels of phenolics, polysaccharides and RNases. Major changes were the inclusion of proteinase K in the extraction medium followed by incubation at 42°C. Integrity and purity were evaluated by denaturing gel electrophoresis and by spectrophotometry (A260/A230 and A260/A280). Total RNA could be successfully used for poly(A)-RNA isolation and cDNA library construction.

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تاریخ انتشار 2005