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Revista Colombiana de Biotecnología
Print version ISSN 0123-3475
Abstract
RUIZ AVILA, Camilo Andrés; GOMEZ VARGAS, Yolanda; BURGOS PAZ, William and TAPIERO, Anibal L.. Protocol for the certification by genetic conformity of clones in rubber propagation gardens (Hevea brasiliensis). Rev. colomb. biotecnol [online]. 2022, vol.24, n.1, pp.56-61. Epub Sep 02, 2022. ISSN 0123-3475. https://doi.org/10.15446/rev.colomb.biote.v24n1.83584.
The polymorphism of 9 regions identified in the DNA of leaf tissue sampled from 12 rubber clones conserved in clonal gardens of AGROSAVIA in Colombia and 25 clones in clonal gardens of origin in Brazil was visualized by capillary electrophoresis after amplification with the fluorolabeled primer microsatellite markers Hmct5, 102, HV 30, 548, HV 15, 416, m574, 103 and 358. Upon the results analysis, a database was consolidated that allows to corroborate the genetical identity by conformity with 37 rubber clones from leaf samples. The established protocol is a methodological approach using end-point PCR towards the amplification by multiplexed capillary electrophoresis of micro-satellite regions and their visualization, reducing costs and optimizing time in the laboratory. Additionally, discrepancies were found between the electrophoretic profile obtained from clone FX 3864 sampled in Colombia with that obtained in Brazil. It is proposed to consider the need to corroborate the identity of the clones reproduced in clonal gardens for their commercialization in Colombia, using sensitive and reproducible methodologies, such as the one standardized in this study.
Keywords : micro-satellite markers; identification; polymorphism.