Preparation of polytene chromosomes in Culex species is very difficult and the available techniques are not always reproducible. Presuming these problems might be due to lack of an appropriate technique to deal with these long and fragile chromosomes invested with ectopic pairings and sticky ends, certain promising methods with several modifications have been tried. During the process, recently, a simple and reproducible method was developed for obtaining good polytene chromosomes from Culex quinquefasciatus. The detailed methodology with precautions and its advantages over other reported techniques are described.