10^8 cells with and without treatment were harvested. Chromatin was sonicated with Tip sonicator (Omni Sonic Ruptor 250 Watt Ultrasonic Homogenizer) for 7 cycles (1 min on (duty cycle 30%, output 20%)) 1 min off) in 2 ml of Lysis buffer 3. The anti-EcR monoclonal antibody (DDA2.7) developed by C. Thummel in D. Hogness laboratory was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52242. 1 ml of chromatin was incubated with 3 μl of antibody (135 μg/ml) overnight. Chromatin and 25 μl of blocked Dynabeads Protein G (Invitrogen, cat. no. 10004D) were combined and incubated at 4° C for additional 2 hrs. 3 ng of material was used for library generation. Library was constructed following instructions of NEBNext® DNA Library Prep Reagent Set for Illumina® (NEB; cat. no. E6000L). Library amplifcation was done using KAPA Library Amp Real Time (KAPA biosystems: cat.no. KK2701).