Revista Chapingo Serie Ciencias Forestales y del Ambiente
Universidad Autónoma Chapingo
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Revista Chapingo Serie Ciencias Forestales y del Ambiente
Volume XVIII, issue 1, January - April 2012


Nadia Guadalupe Sánchez-Coello; Mauricio Luna-Rodríguez; Santiago Mario Vázquez-Torres; Lázaro Rafael Sánchez-Velásquez; Nancy Santana-Buzzy; Pablo Octavio-Aguilar; Lourdes Georgina Iglesias-Andreu

Received: 2011-03-16

Accepted: 2011-11-01

Available online: / pages.123-133


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  • descriptionAbstract

    Most of the cycads contain high concentrations of essential oils, flavonoids, polyphenols, and polysaccharides that interfere with DNA extraction, causing erroneous or no PCR products. The optimization of DNA isolation, employing inter-simple sequence repeats (ISSRs) primers were investigated in Ceratozamia mexicana Brongn., an endangered Mexican cycad. The DNA obtained from fresh-leaf tissues with a modified cetyltrimethylammonium bromide buffer protocol gave a good quality of DNA with no colored pigments and contaminants. The main modification to the CTAB-based DNA extraction protocol was the one hour leaf tissue soaking pre-treatment with a 0.7 M NaCl solution, to facilitate the cell lysis. The DNA extracted was successfully amplified by PCR using six arbitrary ISSR primers. Reproducible amplifiable products were observed in all PCR reactions. Our results show a significant improvement in the DNA quality obtained using low primer concentration (25 pM). 23 strong bands were detected, 9 of which were polymorphic. The results indicated that the optimized protocol for DNA isolation and PCR system is suitable for further work in this specie. This work is the first DNA extraction and ISSR protocols reported for this ornamental and endangered species.

    Keyworks: Ceratozamia mexicana, Cycads, DNA extraction, endangered species, ISSR.
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  • starCite article

    Sánchez-Coello, N. G.,  Luna-Rodríguez, M., Vázquez-Torres, S. M.,  Sánchez-Velásquez, L. R.,  Santana-Buzzy, N., Octavio-Aguilar, P., &  Iglesias-Andreu, L. G. (2012).  OPTIMIZATION OF A PROTOCOL FOR DNA ISOLATION AND ISSR-PCR AMPLIFICATION SYSTEM FOR Ceratozamia mexicana Brongn. (Zamiaceae).. Revista Chapingo Serie Ciencias Forestales y del Ambiente, XVIII(1), 123-133.