Optimization of DNA Extraction for ITS/U4U3 analysis of Rhipsalis baccifera

Tomás Rivas-García; David  Córdova-Pérez; Roberto Carlos  Arredondo-Espinoza; Bernardo  Murillo-Amador; Ramses R.  González-Estrada; Juan José  Reyes-Pérez; Juan Antonio Torres-Rodriguez; Rubí A.  Martínez-Camacho; Jorge Alberto Alejandre-Rosas

doi:10.56890/jpacd.v26i.563

Authors

Tomás Rivas-García CONAHCYT-Universidad Autónoma Chapingo, Texcoco 56230, México.
David Córdova-Pérez Universidad Veracruzana, Orizaba, Veracruz 94340, México
Roberto Carlos Arredondo-Espinoza Université de Caen, Normandie, Caen 14032, France.
Bernardo Murillo-Amador Centro de Investigaciones Biológicas del Noroeste S. C. La Paz, Baja California Sur 23096, México.
Ramses R. González-Estrada Tecnológico Nacional de México/Instituto Tecnológico de Tepic, Tepic 63175, México.
Juan José Reyes-Pérez Universidad Técnica Estatal de Quevedo, Los Ríos 120501, Ecuador.
Juan Antonio Torres-Rodriguez Universidad Técnica Estatal de Quevedo, Los Ríos 120501, Ecuador.
Rubí A. Martínez-Camacho Centro de Investigaciones Biológicas del Noroeste S. C. La Paz, Baja California Sur 23096, México.
Jorge Alberto Alejandre-Rosas Universidad Veracruzana, Orizaba, Veracruz 94340, México.

DOI:

https://doi.org/10.56890/jpacd.v26i.563

Keywords:

Cactaceae, mistletoe cactus, CTAB buffer, DNA yield, molecular markers

Abstract

Genetic analysis of plants relies on high yields of pure DNA. For Rhipsalis baccifera this represents a great challenge since its plant tissue can accumulate large amounts of mucilage, polysaccharides, polyphenols and secondary metabolites, which co-purify with amplifiable DNA. These contaminating compounds lead to a poor yield and prevent acces to PCR-based analysis. These contaminating compounds lead to a poor DNA yield and prevent access to PCR-based analysis. A number of factors, including choice of plant tissue, tissue preparation, and modifications of the extraction buffer, can impact on DNA extraction process. In this study, four different DNA extraction procedures were tested aiming to develop a simple protocol based on CTAB buffer. The obtained results showed that the use of the outer cuticle of old lyophilized tissue allowed reliable results in R. baccifera plants with a good purity ranging from 1.6 to 1.8 and high DNA yield yield ? 500 ng ?L^-1.