In this study, the transesterification of cottonseed oil with methanol was carried out using NaOH. The parameters, such as molar methanol:oil ratio, NaOH %, and temperature effecting the yield and biodiesel quality, were investigated to optimize biodiesel production from cottonseed oil. Below 50 ı C, transesterification of cottonseed oil was not completed at any molar methanol:oil ratios and catalyst percentages in 1 h. In 3:1 methanol:oil ratio, full conversion was not observed. Optimum conversion was obtained at 6:1 methanol:oil ratio and at temperatures between 50 and 60 ı C. Above 60 ı C, some difficulties were encountered in the washing step. Biodiesels that were produced at different molar methanol:oil ratios, NaOH concentrations, and temperatures were tested according to accepted standard methods and the results were compared with standard values.