GSM8465626: PVN, Negative, CUT&RUN; Mus musculus; OTHER
Nuclei from each sample was isolated separately with 1 ml homogenization buffer in a 2-ml dounce homogenizer. Use Pestle B to dounce it 20 times to ensure isolation of nuclei from cells with minimal damage. Transfer the lysis solution and 3 ml homogenization buffer to individual NalgeneTM centrifuge tubes containing 8 ml Sucrose Cushion buffer. Centrifuge for 1 h at 20,000 rpm, 4°C. Carefully remove the middle layer, which contains a lot of white particles. Discard all the supernatants. After discarding the supernatants, nuclei were resuspended in 100 µl of cold nuclear extraction buffer and proceeded to the ConA bead conjugation step following the standard protocol in the CUT&RUN Kit from Epicypher (EpiCypher, #14-1048).