Butterflies are important components of terrestrial biodiversity and are widely used in taxonomic, ecological and molecular studies. Preservation of butterfly specimens is necessary because of their seasonal occurrence and short adult life span; however, preservation conditions may influence DNA recovery. The present study investigated the effect of oven-drying preservation on DNA recovery from five butterfly species, namely Catopsilia pomona, Eurema hecabe, Phalanta phalanta, Ariadne merione and Danaus chrysippus. Specimens were collected from Godapark, Gangapur Road, Nashik, Maharashtra. Fresh and oven-dried specimens were compared. DNA was isolated from 50 mg tissue using the phenol-chloroform method and quantified using the diphenylamine (DPA) colorimetric method. Mean DNA yields from fresh specimens were 96.60, 36.67, 25.00, 46.67 and 93.33 µg, respectively, whereas preserved specimens yielded 26.60, 36.67, 30.00, 23.33 and 43.33 µg, respectively. Reported DNA degradation ranged from 19.3% to 74.0%. The reported t-test values exceeded the critical value for all five species. The study indicates that oven drying can influence DNA recovery and that fresh specimens are preferable when higher DNA yield is required for molecular investigations.