African Journal of Biotechnology Vol. 7 (1), pp. 051-054, 4 January 2008   

ISSN 1684–5315 © 2008 Academic Journals        

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.

*Corresponding author. E-mail: pav20245@cucba.udg.mx. Ph/Fax: (01 33) 36 82 14 63.

Abbreviations: CTAB, hexadecyltrimethylammonium bromide; SDS, sodium dodecyl sulfate; RAPD, random amplified polymorphic DNA; SSR, simple sequence repeats; RFLP, restriction fragment length polymorphism; PVP, polyvinylpyrrolidone.

Accepted 5 November, 2007

Pines are considered to be difficult for DNA extraction. However, from one species to the 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 from Pinus 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.