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Acta Biol Hung ; 64(2): 184-95, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23739887

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.


Subject(s)
DNA, Plant , Genetic Variation , Random Amplified Polymorphic DNA Technique/methods , Saururaceae/chemistry , Saururaceae/genetics , DNA, Plant/chemistry , DNA, Plant/genetics , DNA, Plant/isolation & purification
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