The final volume was adjusted to 100 ml with distilled water. The ground larvae in solution were placed in a boiling water bath for 15 min. The tube is centrifuged at 6,000 rpm to get rid of debris. Larval tissues were centrifuged again at 6,000 rpm for 10 min at 4°C, resuspended in buffer (100 mM Tris-HCl (pH = 8.0), 100 mM EDTA, 250 mM NaCl)