GSM4551965: Adult_CTX_SSTcre_Sun1IP_H3K27ac_HiChIP_rep2; Mus musculus; ChIP-Seq
INTACT-purified nuclei were crosslinked in 1% formaldehyde, followed by the addition of 0.125M glycine to stop crosslinking activity. The nuclei were washed with cold 1XPBS, then lysed once with L1 buffer (50mM HEPES pH7.5, 140mM NaCl, 1mM EDTA, Vip or Sst-expressing nuclei were INTACT-isolated and cross-linked using 1% formaldehyde as described above. HiChIP experiments were performed as previously described (Mumbach et al., 2016) with the following modifications to the protocol: (1) 4 µl of MboI was used to digest chromatin. (2) Sonication step was performed using Covaris M220 (Millitube) PIP 75; Duty factor 5%; CPB 200; Time 2 minutes. (3) For the IP step, 2 µl of anti-H3K27ac antibodies (Abcam ab4729) and 20 µl of Protein A Dynabeads (Thermofisher) were used. For adding Illumina sequencing adapters to samples, we used 0.05 µl of Tn5 (Illumina) to each sample and amplified for 12 cycles using the Nextera DNA sample kit (Illumina).
GEO Accession
GSM4551965