Extraction of DNA from bacteria preserved in the strain bank of the Universidad Francisco de Paula Santander head office Campos Elíseos
Extracción de ADN de bacterias conservadas en el banco de cepas de la Universidad Francisco de Paula Santander sede Campos Elíseos
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Currently, the identification of bacteria is done with classical techniques based on the phenotypic characterization at macroscopic
and microscopic level, however, this method is not reliable and does not allow knowing the true identity of the microorganism
under study. For this reason it is necessary to carry out an identification at the molecular level that allows us to know with
certainty the genus and species. The objective of this work was to standardize a method of DNA extraction for Gram-negative
and Gram-positive bacteria according to the chemical characterization, which allowed obtaining good quality DNA for the
amplification of a specific region of interest. Two extraction methods were evaluated, the Wilson protocol and the Wizzard
extraction kit (Promega), the two methods were differentiated in the quantification by NanoDrop and visualized by means of a
0.8% agarose gel electrophoresis using the Intercalante Gel red. According to Wilson's protocol, only DNA from Gram negative
bacteria of good quality was obtained, and the ratio 260/280 in the nanodrop (1.8-2). While the Wizzard Extraction Kit allowed
to obtain DNA for Gram positive and negative bacteria. Better DNA concentrations were obtained with the Wilson protocol, and
it was not possible to obtain DNA from Gram positive bacteria. For this case, the Wizzard kit was an unlimited method to extract
DNA of any kind, according to the reasons mentioned above, the protocol that reduces costs in the laboratory and is the most
convenient to extract the DNA from Gram negative bacteria is the one proposed by Wilson.
Keywords: Nucleic acids,Bacillus thurigiensis,E. coli,Microorganisms
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