REMOVAL PERFORMANCE PROTOCOL CREOLE CATTLE DNA

Abstract

In order to compare five volumes of commercial reagent applied in the extraction of bovine DNA in plucked hair, random samples of hair from the tail area of 25 Creole cattle breeds Guaymi and Guabala were taken. DNA concentration in nanograms per microliter (ng/ul) was obtained and the purity was determined by the relationship between absorbance at 260 nm and 280 nm (R) for each of the samples. The results of the analysis conducted for treatments showed no significant differences for the value of R. However, for concentration, a significant differences between treatment 5 and 4 (P = 0.019) and treatment 5 and 1 (P = 0.021) were observed. The overall average of R was 1,77 ± 0,22 and concentration was 34,96 ± 7,18 ng/ul. The observed values ​​of R ranging from 1,37 ± 0,60 in treatment 3 and 2,24 ± 0,61 in treatment 2. As to the DNA concentration, the values ​​were in 12,28 ± 12,98 ng/ul in the treatment 4 and 75,21 ± 14,04 ng/ul in the treatment 5. Genomic DNA extraction was achieved in all treatments with similar values ​​of purity. Optimization occurred up to five times extraction capacity of the commercial product and reduced cost per sample; also the sequence of steps of the protocol using a single thermal cycler in the total extraction process was simplified.