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Acta Biologica Hungarica

, Volume 64, Issue 2, pp 184–195 | Cite as

Optimization of DNA Isolation and PCR Protocol for Analysis and Evaluation of Genetic Diversity of the Medicinal Plant, Anemopsis Californica Using RAPD

  • C. Lizette Del-Toro-Sánchez
  • S. Villaseñor-Alvarado
  • Florentina Zurita-Martínez
  • O. A. Castellanos-Hernández
  • Araceli Rodríguez-Sahagún
  • M. Isabel Torres-Morán
  • D. Rojas-Bravo
  • M. Gutiérrez-Lomelí
Open Access
Article

Abstract

Anemopsis californica is a perennial herbaceous plant that has been utilized as a medicinal plant for the treatment of various diseases. The present work was carried out with the objective of optimizing a method of extraction of the genomic DNA of A. californica and a PCR protocol and later to evaluate the existing genetic diversity among the genotypes deriving from different origins. For DNA extraction, we tested four procedures: with the CTA B-2 protocol, we obtained the highest yield (61.5±2.2 µg DNA/g of leaf tissues) and the best quality (A260/280 1.83±0.022). To estimate genetic variability, we utilized the randomly amplified polymorphism DNA (RAPD) technique, employing 20 oligonucleotides, of which only 18 generated reproducible banding patterns, producing 123 polymorphic bands generated, thus obtaining a polymorphism rate of 93.93% among the genotypes analyzed. The Jaccard similarity coefficient generated a variation ranging from 0.325-0.921, indicating a high level of genetic variation among the studied genotypes. An Unweighted pair-group method with arithmetic mean (UPGMA) group analysis indicated six distinct groups. The present optimized method for DNA isolation and RAPD protocol may serve as an efficient tool for further molecular studies.

Keywords

Anemopsis californica DNA isolation RAPD analysis genetic diversity 

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© Akadémiai Kiadó, Budapest 2013

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • C. Lizette Del-Toro-Sánchez
    • 1
  • S. Villaseñor-Alvarado
    • 1
  • Florentina Zurita-Martínez
    • 1
  • O. A. Castellanos-Hernández
    • 1
  • Araceli Rodríguez-Sahagún
    • 1
  • M. Isabel Torres-Morán
    • 2
  • D. Rojas-Bravo
    • 1
  • M. Gutiérrez-Lomelí
    • 1
  1. 1.Department of Medical Sciences and LifeUniversity Center of the Cienega, University of GuadalajaraOcotlánMéxico
  2. 2.University Center of Biological and Agricultural SciencesUniversity of GuadalajaraZapopanMéxico

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