Aglaonema species are herbaceous evergreen and popular cultivated plant for the ornamental plant. In this study there had five different types of Aglaonema species that were analyzed which A. nitidum, A. pumilum, A. simplex, A. cochinchinese and A. flemingianum. The study of DNA isolation and standardization of PCR condition was investigate by using DNA extraction to get genomic DNA and Random Amplified Polymorhic DNA (RAPD) for genetic marker analysis. In using CTAB methods showed a higher yield of DNA quantity and quality. However the young fresh leaf showed the good quality DNA but mature leaves contain higher quantities of polyphenols and polysaccharides, which make it very difficult to isolate DNA of good quality. The Aglaonema DNA was further used for RAPD analysis firstly by using ten random primers and only five random primers used for optimization of annealing temperature. However, the in RAPD analysis showed the negative result and only two from five random primers achieved the optimization of annealing temperature with A06 had the annealing temperature at 37.0°C and while primer D03 with annealing temperature at 35.8°C.