An improved method for high quality RNA isolation from needles of adult maritim...

• Autor(es): Azevedo, Herlânder ; Neto, T. Lino ; Tavares, R. M.
• Data: 2003
• Identificador Persistente: http://hdl.handle.net/1822/2826
• Origem: RepositóriUM - Universidade do Minho
• Assunto(s): cDNA library construction; Maritime pine needles; RNA isolation
• Tipo de Documento: Artigo
• Idioma: Inglês

Descrição

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.