GSM6190686
To prepare nuclei, around 50,000 cells at 500g for 5 min, which was followed by a wash using 50 μL of cold 1× PBS and centrifugation at 500g for 5 min. Cells were lysed using cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2 and 0.1% IGEPAL CA-630). Immediately after lysis, nuclei were spun at 500g for 10 min using a refrigerated centrifuge.  Pools of 5,000-10,000 cortical pyramidal neurons were sorted into PBS. The EZ DNA Methylation-Direct Kit (Zymo Research) was used to perform bisulfite conversion and libraries were prepared with the EpiGnome Methyl-Seq Kit (Illumina).