Optimasi teknik isolasi dan purifikasi DNA yang efisien dan efektif pada kemiri sunan (Reutalis trisperma (Blanco) Airy Shaw) | Optimation of DNA isolation and purification techniques on Reutalis trisperma (Blanco) Airy Shaw

Reutalis trisperma is well known as a potential plant which produces biodiesel and to be used for the conservation as well. The reutalis oil is toxic, therefore it is inedible due to about 50 percent alpha-eleostearat acid content in the oil. As a potential plant, its information in more detail is needed including the DNA analysis. There are many techniques to conduct DNA isolation depending on kind of plants, plant organ, or plant tissue that will be analyzed. The aim of this experiment was to find the effectiveness and efficiency techniques of DNA isolation and purification, so they can reduce cost and time while working in the laboratory. The experiment was conducted at Molecular Biology Laboratory of Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development (BB BIOGEN), Bogor from July to September 2010. Young leaves of reutalis, used as genetic materials were taken from germplasms collection at Pakuwon Experimental Station of Indonesian Spice and Industrial Crops Research Institute (ISICRI), Sukabumi. While some chemicals were used as the other material. The activities were as follows DNA extraction and purification, measurement of DNA concentration, and amplification of DNA. Deletion of resistor enzyme-polysacharide, especially for perennial plant. DNA isolation can be done by breaking down of cell wall, cell membrane, and nuclear membrane. The results showed that conscientiousness of DNA isolation and purification denoted an important step to obtain clean and contaminant free of DNA, so the banding patterns were clear. In this technique did not use polypinilpolypirolidone (PYPP) and mercapto-ethanol such as antioxidant, liquid nitrogen, neither over night storage of leaf extraction before used for purification which is often used for perennial plant. In addition the results showed that band pattern of DNA was very thick and clear, therefore this technique can be applied for DNA isolation on Reutalis trisperma.