title: GSM8724787: HA ChIP, HA-Lola-I, rep1; Drosophila melanogaster; ChIP-Seq
library_construction_protocol: 30 third-instar larval CNSs per biological replicate (two biological replicates per sample) were dissected in ice-cold PBS. Chromatin was prepared as described in (Gambetta, Oktaba and Müller, 2009) with the following modifications: inverted larval cuticles were cross-linked for 15 minutes at room temperature in 1.8% (v/v) paraformaldehyde, and dissected brains were sonicated in 120 µl of previously described RIPA140 buffer in AFA microtubes in a Covaris S220 sonicator (Covaris) for 5 minutes with a peak incident power of 140W, a duty cycle of 5% and 200 cycles per burst. Sonicated chromatin was centrifuged to pellet insoluble material and snap-frozen.  ChIP was performed with rabbit polyclonal anti-HA antibody (Abcam Ab9110). ChIP was performed as previously described in (Gambetta, Oktaba and Müller, 2009) except that Protein A and G Dynabeads were used. ChIP-seq libraries were prepared using the NEBNext Ultra II DNA Library Prep kit for Illumina. An equimolar pool of multiplexed ChIP-seq libraries was sequenced 150 bp paired-end. Standard ChIP-seq libraries were prepared with the NEBNext Ultra II DNA Library Prep kit for Illumina.