| dc.description.abstract | Deoxyribonucleic Acid (DNA) is smallest part (molecule) of a cell in the form of a long chain that carries genetic information and controlling all cellular functional in all life form. The DNA can be obtained by extracting tissue or fresh whole blood of animal. There are several methods of DNA extraction, one of which is phenol-choroform-isoamyl alcohol (PCI) that used in this research. The aim of this research was to determine the best combination of fresh blood and buffer lysis with two differences of spinning speed on DNA quantity and quality. The fresh whole blood of Friesian Holstein (FH) dairy cattle was collected using a vacuum tube (vacutainer) containing 15% EDTA with a 18G needle to suck the blood of tail vena with the amount of ±5 ml per head. A 3x2 factorial completely random design was used. First factor was three combinations of fresh whole blood and buffer lysis volume (μl), i.e., 100 : 800, 200 : 700, and 300 : 600. Second factor was spinning speeds, i.e., 10.000 rpm and 12.000 rpm. There were 6 combination treatments used in this research which replicated 4 times of each treatment. Data of DNA concentration was collected by a GeneQuant DNA calculator at the 260 nm and 280 nm wave length of optical density (OD). The quality of DNA was obtained by running the DNA through electrophoresis with 1% gel agarose and electricity flow of 100 voltage for an hour. The DNA purity was examined at OD 260/280 ratios. DNA concentration and purity were analyzed by using ANOVA, the differences between theatments were tested by tukey test at the level of 5% (p<0.05). The result showed that there was no significant effect of combination of fresh blood and buffer lysis at both spinning speeds (p>0.05). The highest DNA concentration was obtained from the treatment of ratio of 300 μl blood and 600 μl buffer lysis. The DNA concentration tended to increase with the increase of blood volume. The DNA purity was obtained at the ratio of 1.3 (260/280) meaning that the DNA was a slighty contaminated by the chemical used. A ratio of 1.8 is the optimal purity of DNA. The DNA quality linearly correlated to DNA concentration and showed a single band. Keywords: blood, DNA extraction, phenol-choroform-isoamyl alcohol (PCI), buffer lysis, spinning speed | id |
