GSM6593502: Wild_type_H3K27me3_CST_rep_6; Mus musculus; OTHER
Nuclei were purified from wild-type, MECP2-TG1, or Mecp2-knockout hippocampi according to Mo et al. Purified nuclei were bound to concavalin A beads and incubated with the indicated antibody. Next, nuclei were incubated with pAG-MNase to cleave around antibody-bound DNA regions. Release fragments were collected and used for library prep. CUT&RUN libraries were prepared using NEB Next II DNA Ultra Kit (NEB #E7645S) with NEB Illumina Unique Combinatorial Dual Index Kit (NEB # E6442S). 25 uL CUT&RUN fragments were used to prepare libraries for MeCP2 and IgG samples, 6 ng of input was used for H3K27me3 samples. Libraries were amplified for 14 cycles and fragments. Post amplified libraries were selected for ~300bp in length using SPRI beads from Beckman Coulter. Libraries size distributions were measured using Agilent TapeStation HS DNA 1000 (Agilent #5067) and concentrations were determined using the KAPA library quantification kit (Roche #07960140001).