GSM4498303: NURF301_CUTandRUN; Drosophila melanogaster; OTHER
CUT&RUN was performed as previously described in Skene et al. 2017 (PMID: 28079019). Antibodies to GAF (lab made; 1:10) and NURF301 (Novus Biologicals Cat. No. 40360002; 1:10) were incubated with 10e6 permeabilized Drosophila S2 cells bound to concanavalinA coated beads, and proteinA-MNase cleavage was initiated by incubating on ice with calcium for 30 min. Cleaved fragments were recovered by phenol:chloroform extraction followed by ethanol precipitation. Library preparation was completed using enzymes from NEB (T4 PNK, T4 DNA Polymerase, and Klenow fragment for end-repair; Klenow exo- for A-tailing; and T4 DNA ligase for adapter ligation) and lab-made Illumina forked TruSeq adapters synthesized by IDT. Libraries were amplified for 15 PCR cycles.
GEO Accession
GSM4498303