Extraction of genomic DNA from solid tissues of teleostei fish

Nilton Garcia Marengoni, Marcia Regina Fragoso Machado, Eliane Gasparino

Abstract


The object of this work was to extract the genomic DNA of solid tissue from Nile tilapia (Oreochromis niloticus), pacu (Piaractus mesopotamicus), piau (Leporinus sp.) and curimba (Prochilodus lineatus), using the methods Proteinase K (PK) and Cetyltrimethylammiun Bromide (CTAB). The DNA extracted from samples of liver, kidney, tail fin, heart, muscle and gills were quantified in spectrophotometer to determine the concentration and purity through the ratio A260nm/A280nm. The data were statistically analyzed and there were no significant effect of interaction between species and tissues about the purity and concentration of the DNA obtained with CTAB, but for the PK there were interaction about the concentration. Using the CTAB method was verified that the mean quantity of DNA in the curimba kidney was significantly (p < 0.05) lower (106.98 ?g/mL) than the value observed in pacu, whereas, did not differ from the variables found in piau (497.20 ?g/mL) and tilapia (234.50 ?g/mL). For the PK method, the mean quantity of DNA using the muscle of Nile tilapia presented the lowest value (117.35 ?g/mL) with reference to the other tissues and analyzed species (P < 0.05). The ratio A260nm/A280nm changed from 1.7 to 2.0 and 1.6 to 2.1 for the PK and CTAB methods, respectively. It may be concluded that the DNA purity was satisfactory for the different tissues on the four species of fish.


Keywords


Leporinus sp; Oreochromis niloticus; Piaractus mesopotamicus; Prochilodus lineatus; DNA.



DOI: http://dx.doi.org/10.5433/1679-0359.2006v27n1p99

Semina: Ciênc. Agrár.
Londrina - PR
E-ISSN 1679-0359
DOI: 10.5433/1679-0359
E-mail: semina.agrarias@uel.br
Este obra está licenciado com uma Licença Creative Commons Atribuição-NãoComercial 4.0 Internacional