African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12193

Full Length Research Paper

High yield and high quality DNA from vegetative and sexual tissues of Mexican white pine (Pinus ayacahuite)

Verónica Palomera-Avalos*, Patricia Castro-Félix and Alma R. Villalobos-Arámbula
Departamento de Biología Celular y Molecular, Universidad de Guadalajara, 45110, Zapopan, Jalisco, México.
Email: [email protected]

  •  Published: 04 January 2007

Abstract

 

Pines are considered to be difficult for DNA extraction. However, from one species tothe other there is variation in phenolic profiles and seed size that might affect final DNA yields and quality. Two DNA extraction protocols (CTAB and SDS based) were compared for their ability to produce DNA on leaves, gametophyte and embryo fromPinus ayacahuite, a pine species with a large seed (8 - 18 mm). The DNA obtained from both procedures was quantified and tested by PCR. The CTAB protocol provided higher DNA yields from vegetative tissue and embryo than the SDS method. Embryos (2n) and gametophytes (n) proved to be very good sources of DNA and the DNA isolated was suitable for PCR-RAPD and SSR markers. This paper reports the results and describes the modified CTAB protocol.

 

Key words: CTAB, DNA isolation, Pinus ayacahuite, SDS, seed tissue, vegetative tissue.