| dc.description.abstract | Deoxyribonucleic acid is genetic materials that inherited from one generation to next generation. DNA can be obtained from all body parts of animal, such as tissue and blood. Blood is one of DNA sources that is usually used because of its simple collection. There are several methods to extract DNA, one of which is a high salt method that used in this experiment. The aim of this experiment was to determine the optimal combination of whole blood and buffer lysis ratio at different centrifugation speed (rpm) to obtain optimal DNA products with high quality. Fresh whole blood of Friesian Holstein (FH) dairy cattle was collected by using a vacuum tube containing 15% EDTA with a 18G needle to suck the blood from tail vena with amount of ± 9 ml per head. The whole blood was collected from four FH cows. A 3x2 factorial completely random design was used in this experiment. The first factor was three combinations of whole blood and buffer lysis volume (ml), which was 1:2, 1:1, and 2:1. The second factor was centrifugation speed, i.e. 2,500 rpm and 3,500 rpm. Therefore, there were six combination treatments with four replications of each combination treatment. Data of DNA concentration and purity was observed from Genequant DNA calculator at 260 Å and 280 Å wave length. The quality of DNA was obtained from running DNA through electrophorosis of 0.8% gel agarose and electricity flow of 100 voltages for an hour. The DNA concentration and purity were analyzed by ANOVA. The differences among treatments were tested by Tukey test at = 0.05. The result showed that there was no significant effect of whole blood and buffer lysis combination on DNA concentration and purity. Centrifugation speed had a significant effect on DNA concentration (P<0.05) but did not affect their purity. DNA quality was good because it had a single band. Keywords: blood, DNA, centrifugation speed, lysis buffer, high salt method, cow | id |
